1. the preservation number of the strain of Bacillus subtilis D8 is CGMCC NO. 19552.

2. Use of Bacillus subtilis d8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same of the claims in oil production;

use of Bacillus subtilis d8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in the preparation of petroleum products.

3. Use of Bacillus subtilis D8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in petroleum microbial oil recovery.

Use of Bacillus subtilis d8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in the preparation of microbial oil recovery products.

4. Use of Bacillus subtilis D8 or a bacterial suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial inoculum containing the same in the emulsification of crude oil.

Use of Bacillus subtilis d8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in the preparation of a crude oil emulsified product according to the claims.

5. Use of Bacillus subtilis D8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in petroleum degradation.

Use of Bacillus subtilis d8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in the preparation of petroleum degradation products.

6. Use of Bacillus subtilis D8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in petroleum pollution treatment.

Or, the use of Bacillus subtilis D8 or its bacterial suspension or its culture solution or its fermentation product or microbial inoculum containing it in the preparation of products for treating petroleum pollution.

7. Use of Bacillus subtilis D8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in petroleum pollution remediation.

Or, the use of Bacillus subtilis D8 or a bacterial suspension or a culture solution or a fermentation product thereof or a microbial inoculum containing the same in the preparation of products for repairing petroleum pollution.

8. A product, wherein the active ingredient is Bacillus subtilis.d8 or its bacterial suspension or its culture solution or its fermentation product or a microbial inoculum containing it according to claim 1;

the new product has any one of the following functions 1) to 6):

1) oil exploitation;

2) microbial oil recovery;

3) emulsifying crude oil;

4) degrading petroleum;

5) treating petroleum pollution;

6) and (4) repairing petroleum pollution.

9. A method of microbial oil recovery comprising the steps of: in the process of oil recovery, Bacillus subtilis.d8 or a suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial inoculum containing the same as claimed in claim 1 is added.

10. A method for treating and/or remediating petroleum pollution, comprising the steps of: treating a petroleum pollutant with Bacillus subtilis D8 or a suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial inoculum containing the same as defined in claim 1.

Background art:

petroleum is a non-renewable resource, and with the increasing shortage of petroleum resources and the increasing exploration cost, about 60 to 70 percent of crude oil in the stratum can not be extracted after primary oil extraction and secondary oil extraction. In recent years, tertiary Oil Recovery methods represented by Microbial Enhanced Oil Recovery (MEOR) technology have been receiving wide attention. The technology utilizes beneficial activities and metabolites of microorganisms to improve the crude oil recovery ratio, and is a comprehensive technology following traditional three-recovery methods such as thermal flooding, chemical flooding, polymer flooding and the like.

The microbial oil production technology is a new technology with high technological content and rapid development, is the pioneering application of modern biology and technology in the field of oil production engineering, and shows strong vitality for oil reservoirs with high water content and nearly exhausted oil reservoirs. The microbial oil recovery technology mainly comprises two types (Ronald M. atlas. Petroleum Microbiology [ M ]. New york. Macmill press). one type is that microbial metabolites such as biopolymer and biological emulsifier are used as oil field chemical agents to displace oil, and is called microbial aboveground fermentation enhanced recovery process, namely a biological process method. At present, the technology tends to mature at home and abroad: the other is to increase the recovery ratio by using microorganisms and metabolites thereof, mainly by using the underground fermentation of the microorganisms and by using the vitality of the microorganisms inherent in the oil layer, which is called the method for increasing the recovery ratio by using the underground fermentation of the microorganisms.

The microorganism takes crude oil hydrocarbon as a carbon source, metabolizes biological emulsifiers (such as glycolipid, phospholipid, lipopeptide, fatty acid, short peptide and the like) which are small molecules with low molecular weight, and the biological emulsifiers can obviously reduce the tension of the water interface of the oil ー, thereby being beneficial to oil-water emulsification, leading the alkane to be in colloidal dispersion, increasing the contact between the alkane and cells and improving the degradation efficiency of the microorganism on the hydrocarbon.

The separation of microorganisms for degrading petroleum and producing high-yield effective metabolites is the basis of the microbial enhanced oil recovery technology, and the quality of strains is the key of microbial oil recovery. Among the petroleum microorganisms separated at present, the thermophilic salt-resistant bacteria capable of degrading crude oil under aerobic or anaerobic conditions are similar to the bacteria capable of producing biological emulsifiers. The search for suitable microorganisms is still an important task in the art today.

The petroleum pollution is widely distributed, and relates to various industries such as oil extraction, oil refining, chemical engineering, machinery and the like, and various polluted environments such as petroleum leakage and the like. The petroleum hydrocarbon components are complex and diverse, and have strong hydrophobicity, so that the petroleum hydrocarbon components are difficult to be contacted, degraded and utilized by common microorganisms. The microbial oil recovery technology is to inject nutrients or microbes into the stratum and utilize the growth and metabolism activity of the microbes in the oil reservoir to raise the yield and recovery rate of crude oil.

Disclosure of Invention

One purpose of the invention is to provide a strain of Bacillus subtilis D8

The preservation number of Bacillus subtilis D8 provided by the invention is CGMCC NO.19552, which is classified and named as Bacillus subtilis and is preserved in China general microbiological culture Collection center (CGMCC for short, address: No. 3 Siro No.1 of Beijing Indoron Suzhongchen province, institute of microbiology, China academy of sciences, postal code 100101) 4 months and 3 days in 2020

Another object of the present invention is to provide a novel use of Bacillus subtilis.d8 or a suspension thereof or a culture solution thereof or a fermentation product thereof or a microbial agent containing the same.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum recovery.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of petroleum products.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in microbial oil recovery.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the fermentation product thereof in preparation of microbial oil recovery products.

The invention provides application of Bacillus subtilis D8 or suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the same in crude oil emulsification,

the invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of crude oil emulsified products.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum degradation.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of petroleum degradation products.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum pollution treatment.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of products for treating petroleum pollution.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in petroleum pollution remediation.

The invention provides application of Bacillus subtilis D8 or suspension thereof, or culture solution thereof, or fermentation product thereof, or microbial inoculum containing the same in preparation of products for repairing petroleum pollution.

It is another object of the invention to provide a product

The active ingredient of the product provided by the invention is the Bacillus subtilis.D8 or the suspension thereof or the culture solution thereof or the fermentation product thereof or the microbial inoculum containing the fermentation product thereof;

the product has any one of the following functions:

1) oil recovery;

2) microbial oil recovery;

3) emulsifying crude oil;

4) degrading petroleum;

5) treating petroleum pollution;

6) and (4) repairing petroleum pollution.

It is yet another object of the present invention to provide a method for microbial oil recovery.

The invention also provides a microbial oil recovery method, which comprises the following steps: adding the Bacillus subtilis D8 or suspension thereof or culture solution thereof or fermentation product thereof or microbial inoculum containing the Bacillus subtilis D8 or the suspension thereof or the fermentation product thereof in the oil extraction process.

It is a final object of the present invention to provide a method for remediation and/or rehabilitation of petroleum pollution. The method for treating and/or repairing petroleum pollution provided by the invention comprises the following steps: and (3) treating the petroleum pollutants by using the Bacillus subtilis D8 or the suspension thereof, the culture solution thereof, the fermentation product thereof or the microbial inoculum containing the fermentation product.

In the above application or product or method, the preparation method is as follows: inoculating Bacillus subtilis D8 into a culture medium for culture to obtain the microbial inoculum. The concentration of the bacteria in the microbial inoculum is not less than 10⁸cfu/ml

The invention provides a microbial oil-producing strain, which is classified and named as Bacillus subtilis D8, and is preserved in China general microbiological culture Collection center (CGMCC) at 4-3.2020, with the preservation number of CGMCC NO. 19552. Experiments prove that: the microbial oil recovery bacterium can emulsify crude oil, can be used for microbial oil recovery and oil recovery rate improvement, can also be used for treatment and restoration of petroleum pollution, and has good application prospect.

Drawings

FIG. 1 is a graph of the displacement results of a model.

Deposit description

Latin name: bacillus subtilis

The strain number is as follows: d8

The preservation organization: the China Committee for culture Collection of microorganisms general microbiological culture Collection center (CGMCC) is abbreviated as: CGMCC (China general microbiological culture Collection center)

Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district

The preservation date is as follows: year 2020, 4 and 3

Deposit center stamp accession number: CGMCC NO.19552

Detailed Description

The following examples are provided to facilitate understanding of the present invention, but are not intended to limit the present invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates, the average.

Examples 1, 1 isolation, identification and preservation of strains

Isolation of D8 Strain

An oil-water mixture is obtained from oil well produced liquid of a victory oil field of eastern Shandong in 2019, and the inventor separates a strain from the oil well produced liquid and names the strain D8.

II, identification of the strains

1. Morphological characterization of strains

Morphological characteristics of the strain: the diameter of a colony is 2-3mm in one day, the bacterial strain forms a transparent convex lens-shaped round colony on a culture medium, and the colony is picked up by an inoculating needle to have a wire drawing phenomenon.

2. Physiological and biochemical identification of strains

The cell morphology characteristics of the Bacillus subtilis D8, CGMCC NO.19552 provided by the invention are as follows: the thallus is gram-positive, and under the microscope, the cell is rod-shaped, terminal spore-forming and short rod-shaped. The growth temperature range of the strain is 35-75 ℃, the most suitable growth temperature is 60 ℃, the growth pH range is 6-11, the NaCl tolerance is 0-18%, and a puncture experiment shows that D8 is facultative anaerobe;

part of the physiological and biochemical characteristics of the Bacillus subtilis D8 and CGMCC NO.19552 of the invention are shown in Table 1:

TABLE 1 physiological and biochemical characteristics of D8 strains

3. Molecular characterization of strains

Inoculating the activated strain into liquid LB culture medium (water as solvent, peptone and yeast powder as solute and concentration respectively), and shake-culturing for 24 h. And (3) centrifuging 1mL of fresh culture solution, collecting thalli in a centrifuge tube, and extracting DNA by using a DNA extraction kit. After electrophoretic detection, universal primers are used for amplification. After electrophoresis detection of the product, determining the gene sequence, wherein the specific sequence is as follows: 16SrDNA sequence determination results and gyrB gene sequence determination results.

Third, preservation of the Strain

The classified name of the strain is Bacillus subtilis, which has been preserved in China general microbiological culture Collection center (CGMCC for short, address: No. 3 of Beijing institute of microbiology, Japan institute of sciences, Japan, postal code 100101) in China Committee for culture Collection of microorganisms, 4.3.2020, 4.s.and 3.s..

Example 2 Performance experiment of emulsified crude oil by Bacillus subtilis D8 CGMCC NO.19552 Strain of the invention under aerobic conditions

1. Test strains

The preservation number of the strain of the Bacillus subtilis separated in the embodiment 1 of the invention is CGMCC NO. 19552.

2. Experimental methods

The test strain grows in inorganic salt emulsion culture medium with molasses as carbon source, and inorganic salt emulsion culture medium D8 bacterial liquid, (NH)₄)₂SO₄、Na₂HPO₄、NaNO₃Peptone, yeast extract and molasses, the concentration of which in the bacteria-containing water is respectively 5% and 1g/L (NH)₄)₂SO₄、 1g/LNa₂HPO₄、3g/LNaNO₃2g/L peptone, 2g/L yeast extract and 2g/L molasses.

The culture was carried out at 65 ℃ for 2 days with shaking (120 rpm). Taking fermentation liquor at different time points, diluting the fermentation liquor by 20 times with sterile water, taking fermentation stock solution and diluent to emulsify the crude oil, measuring an emulsification index, measuring the surface tension of the fermentation liquor by using a surface tension meter, and observing the emulsification condition of the fermentation liquor on the crude oil.

The method for measuring the emulsification index comprises the following steps: taking two graduated test tubes, respectively adding 3ml of diesel oil and 3ml of crude oil, adding 7ml of fermentation liquor into each test tube, violently oscillating for 1 minute, standing at room temperature for 24 hours, measuring, dividing the height of an emulsion layer by the total height of an organic phase, and multiplying by 100 percent to obtain EI; the emulsion is also considered stable if the EI is > 50%.

Surface tension measurement method: 2ml of the fermentation liquid is taken, centrifuged at 12000rpm/min for 5 minutes, 1ml of the supernatant is taken, and the surface tension of the fermentation liquid is measured by a surface tension meter.

3. Results of the experiment

The experimental result shows that the test strain can well emulsify the crude oil, so that the crude oil particles become fine and are fully dispersed in water, such as an ink juice shape, and the crude oil particles are mutually soluble with the water.

Inorganic salt culture medium D8 bacterial liquid, (NH)₄)₂SO₄、Na₂HPO₄、NaNO₃Peptone, yeast extract and molasses, the concentration of which in the bacteria-containing water is respectively 5% and 1g/L (NH)₄)₂SO₄、1g/LNa₂HPO₄、3g/LNaNO₃2g/L peptone, 2g/L yeast extract and 2g/L molasses.

The test strain can generate a surfactant in an inorganic salt emulsification medium taking molasses as a carbon source, so that the surface tension of fermentation liquor is remarkably reduced, the surface tension of the fermentation liquor is reduced from 67.2mN/m to 25.6mN/m, the reduction amplitude exceeds 50%, the emulsification of crude oil is well promoted, and the emulsification index EI-24 can reach 73% at most.

The test strain can achieve good emulsification effect on crude oil only in 40 hours under the aerobic condition of 65 ℃.

Example 3 performance test of viscous substance production under facultative anaerobic conditions,

inorganic salt culture medium D8 bacterial liquid with cane sugar as carbon source, (NH)₄)₂SO₄、Na₂HPO₄、 NaNO₃Peptone, yeast extract and sucrose to make their concentrations in bacteria-containing water respectively 5% and 1g/L (NH)₄)₂SO₄、1g/LNa₂HPO₄、3g/LNaNO₃2g/L peptone, 2g/L yeast extract and 2g/L sucrose.

The test strain grows in an inorganic salt culture medium taking cane sugar as a carbon source under a facultative anaerobic condition, and the culture solution is put into an anaerobic bottle and is statically cultured for 2 days at 60 ℃. It can be observed that a layer of flocculent precipitate appears at the bottom of the fermentation broth, the flocculent precipitate disappears after shaking, the viscosity of the fermentation broth is tested by a Brookfield viscometer, the viscosity is 6.2mPa.S, and the viscosity of the corresponding blank sterile fermentation broth is 0.8 mPa.S.

The test strain produces viscous substances under the facultative anaerobic condition of 60 ℃ and under the high temperature and specific conditions, and the extracellular polysaccharide produced by the test strain is dissolved in the injected water, so that the swept efficiency of the injected water is enhanced, and the crude oil recovery rate is improved.

Example 4 core Profile control physical simulation experiment

The preparation method of the bacteria-containing water comprises the following steps: inoculating the activated strain into a liquid LB culture medium, and statically culturing for 24H to obtain a bacterial liquid. Adding D8 bacterial liquid and (NH) into the injected water₄)₂SO₄、 Na₂HPO₄、NaNO₃Peptone, yeast extract and sucrose to make their concentrations in bacteria-containing water respectively 5% and 1g/L (NH)₄)₂SO₄、1g/LNa₂HPO₄、3g/LNaNO₃2g/L peptone, 2g/L yeast extract and 2g/L sucrose.

The core simulation experiment adopts an outcrop homogeneous core, the permeability of the core is measured by gas, the core is vacuumized for more than 8 hours, the core is saturated by formation water, the Pore Volume (PV) is calculated, water injection displacement is carried out until the pressure is stable, then 0.1PV fermentation liquor is injected, the inlet and outlet valves of the core are closed, the process water drive is reinstalled after the core is placed in a 55 ℃ incubator for 3 days, and the pressure change before and after injection profile control is measured.

Due to the blocking effect of thalli and polysaccharide generated by the thalli on the core pores, the core injection pressure is increased to 0.4MPa from 0.1MPa before profile control, the core permeability is reduced to 0.12um2 from 0.556um2, the permeability reduction rate is 80%, and the experiment result proves that extracellular polysaccharide generated by the strain D8 has an effective profile control and blocking effect on the core pores.

The strain D8 utilizes nutrition to generate extracellular polysaccharide to form a biofilm in a rock core, and can utilize residual oil in the rock core to generate surface active substances while blocking rock pores to increase pressure and adjust permeability, so that crude oil is emulsified, the fluidity of the crude oil is increased, the recovery ratio is improved, and the microbial effect can be considered as the microbial profile control effect.

Example 5 application of strains in microbial oil-displacing technology

In the embodiment, the petroleum is collected from Shandong Shengli oil field in 2019, 8 months and 10 days, and the viscosity of the crude oil is 500mPa.S

In the embodiment, the oil displacement effect of the microbial strains is evaluated through an indoor physical model oil displacement experiment. The specific experimental method is as follows:

the high-pressure model pipe is filled with quartz sand with different meshes to be used as a sand filling model pipe. And (4) adopting a permeability measuring device to measure the permeability of the model pipe. The formation water is saturated by vacuum pumping. Calculating the porosity; saturating crude oil in a rock core, creating bound water, and measuring the original oil saturation; and aged at reservoir temperature 60 ℃ for 3 days.

Connecting the injection process, injecting water to drive oil, stopping water drive until the water content of the outlet produced liquid reaches more than 98%, and calculating the recovery ratio (namely the water drive recovery ratio).

Injecting 0.5PV bacteria-containing water according to the pore volume of the core (calculating the recovery ratio after injection is completed, namely the recovery ratio in the microbial injection process), and culturing for 10 days at the oil reservoir temperature of 60 ℃.

The preparation method of the bacteria-containing water comprises the following steps: inoculating the activated strain into a liquid LB culture medium, and performing shaking culture for 24H to obtain a bacterial liquid. Adding D8 bacterial liquid and (NH) into the injected water₄)₂SO₄、 Na₂HPO₄、NaNO₃Peptone and yeast extract, wherein the concentration of the peptone and the yeast extract in the bacteria-containing water is respectively 5% and 1g/L (NH)₄)₂SO₄、1g/LNa₂HPO₄、3g/LNaNO₃2g/L peptone and 2g/L yeast extract.

And after 10 days, performing secondary water injection to drive oil, stopping water drive until the outlet produced liquid reaches the limit water content, and calculating the recovery ratio (namely the recovery ratio of the subsequent water drive under the action of the microorganisms). The physical model displacement result curve is shown in figure 1.

The recovery ratios in different stages of water flooding, microbial injection, subsequent water flooding under the action of microbes and the like are compared, and the results are shown in table 2. The indoor physical model displacement of reservoir oil shows that the injection pressure of the water displacement is increased again, which shows that the crude oil forms emulsion, the water content is reduced by 15 percent, the recovery rate is improved by 13 percent, and the D8 strain can be used for improving the recovery rate.

TABLE 2 oil displacement result table for indoor physical model

Moulding process	Primary water drive recovery ratio%	Enhanced recovery ratio%
			Water drive	43.3％
Microbial flooding	67.31％	13％

1. 16S rDNA sequence determination results:

ATACATGCAAGTCGAGCGGACAGATGGGAGCTTGCTCCCTGATGTTAGCGGCGGACGGGTGAGTAACACGTGGGTAACCTGCCTGTAAGACTGGGATAACTCCGGGAAACCGGGGCTAATACCGGATGGTTGTTTGAACCGCATGGTTCAAACATAAAAGGTGGCTTCGGCTACCACTTACAGATGGACCCGCGGCGCATTAGCTAGTTGGTGAGGTAATGGCTCACCAAGGCAACGATGCGTAGCCGACCTGAGAGGGTGATCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCGCAATGGACGAAAGTCTGACGGAGCAACGCCGCGTGAGTGATGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTAGGGAAGAACAAGTACCGTTCGAATAGGGCGGTACCTTGACGGTACCTAACCAGAAAGCCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTCGCAGGCGGTTTCTTAAGTCTGATGTGAAAGCCCCCGGCTCAACCGGGGAGGGTCATTGGAAACTGGGGAACTTGAGTGCAGAAGAGGAGAGTGGAATTCCACGTGTAGCGGTGAAATGCGTAGAGATGTGGAGGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAGTGCTAAGTGTTAGGGGGTTTCCGCCCCTTAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGGTCGCAAGACTGAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAGGTCTTGACATCCTCTGACAATCCTAGAGATAGGACGTCCCCTTCGGGGGCAGAGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGATCTTAGTTGCCAGCATTCAGTTGGGCACTCTAAGGTGACTGCCGGTGACAAACCGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACCTGGGCTACACACGTGCTACAATGGACAGAACAAAGGGCAGCGAAACCGCGAGGTTAAGCCAATCCCACAAATCTGTTCTCAGTTCGGATCGCAGTCTGCAACTCGACTGCGTGAAGCTGGAATCGCTAGTAATCGCGGATCAGCATGCCGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACACCACGAGAGTTTGTAACACCCGAAGTCGGTGAGGTAACCTTTTAGGAGCCAGCCGCCGAAGGTG

2. determination of gyrB Gene sequence:

TATAAACGCGGAGTTCCGGTTACAGACCTTGAAATCATTGGCGAAACGGATCATACAGGAACGACGACACATTTTGTCCCGGACCCTGAAATTTTCTCGGAAACAACCGAGTATGATTACGATCTGCTTGCCAACCGCGTGCGTGAATTAGCCTTTTTAACAAAGGGCGTAAACATCACGATTGAAGATAAACGTGAAGGACAAGAGCGCAAAAATGAATACCATTACGAAGGCGGAATTAAAAGTTATGTAGAGTATTTAAACCGCTCTAAAGAGGTTGTCCATGAAGAGCCGATTTACATTGAAGGCGAAAAGGACGGCATTACGGTTGAAGTGGCTTTGCAATACAATGACAGCTACACAAGCAACATTTACTCGTTTACAAACAACATTAACACGTACGAAGGCGGTACCCATGAAGCTGGCTTCAAAACGGGCCTGACTCGTGTTATCAACGATTACGCCAGAAAAAAAGGGCTTATTAAAGAAAATGATCCAAACCTAAGCGGAGATGACGTAAGGGAAGGGCTGACAGCGATTATTTCAATCAAACACCCTGATCCGCAGTTTGAGGGCCAAACGAAAACAAAGCTGGGCAACTCAGAAGCACGGACGATCACCGATACGTTATTTTCTACGGCGATGGAAACATTTATGCTGGAAAATCCAGATGCAGCCAAAAAAATTGTCGATAAAGGCTTAATGGCGGCAAGAGCAAGAATGGCTGCGAAAAAAGCGCGTGAACTAACACGCCGTAAGAGTGCTTTGGAAATTTCAAACTTGCCCGGTAAGTTAGCGGACTGCTCTTCAAAAGATCCGAGC