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Transgene enterobacter cloacae CEI-a and its construction method and use

A kind of Enterobacter cloacae, transgenic technology, applied in biological field

Inactive Publication Date: 2011-08-10
GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The question is how to introduce serine protease inhibitors into the RIFA population and play a role, which is still an unresolved topic

Method used

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  • Transgene enterobacter cloacae CEI-a and its construction method and use
  • Transgene enterobacter cloacae CEI-a and its construction method and use
  • Transgene enterobacter cloacae CEI-a and its construction method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0025] 1) Extract the total RNA of soybean Glycine max L seeds, reverse transcription and clone the serine protease inhibitor Tia gene

[0026] The primer design part is based on the cloned Tia gene sequence (GenBank accession AB112031) abroad and the multi-cloning site situation of the transposable plasmid pMini-Tn5, using Primer Premier5.0 and DNAStar software to assist in the design of primers. A Not I restriction site was introduced at the 5' end of the upstream and downstream primers, and Not I had no cutting site inside the cloned gene. Wherein, the upstream primer introduces a section of SD sequence rich in purine bases (in the box) at a position 7 bases away from the ATG initiation codon of the target gene sequence. The designed primer sequences are as follows:

[0027] Tia-for-b5′- GCGGCCGC CCCAAAAATGAAGAGCAC-3′

[0028] Tia-rev5′-A GCGGCCGC GTCTCACTCACTGCGAGAAAG-3′

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Abstract

The present invention provides transgenic enterobacter cloacae (Enterobacter cloacae) GEI-a, and the construction method and the purpose thereof. The bacterial strain utilizes a Tn5 transposon to insert a soybean serine proteinase inhibitor gene into the genome DNA of the enterobacter cloacae, to be used to express engineering strain which inhibits the insect intestinal tract trypsin. The strain preserves in the China center for type culture collection, and the serial number is CCTCC M 207071. Fire ant symbiotic bacteria for expressing an exogenous gene is used as a bait to feed the fire ants, recombinant bacterium reproduces in the intestine cavity of the fire ant, and the albumen which can cause the death of the fire ant is produced. The present invention is the secure and effective method to incessantly control the fire ants.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a transgenic Enterobacter cloacae strain, its construction method and its application. Background technique [0002] Plant protease inhibitors (Protein inhibitor, PI for short) are a class of low-molecular-weight polypeptides or proteins that widely exist in the plant kingdom, and have inhibitory effects on insect proteases. When protease inhibitors are fed by insects and accumulate in the body, it can bind to the active site or allosteric site of insect protease, inhibit the catalytic activity of protease, disrupt the normal metabolism of insects, cause insect indigestion, and cause a lack of essential amino acids. Thereby, the growth and development of insects are hindered, and even die. Thus these inhibitors have great potential in pest control [Bowler1990 Defense-related proteins in higher plants. Annual Review of Biochemistry, 59: 873-907; Liu Wuge et al. 2000 Prote...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29A01N63/02A01P7/04C12N1/21C12R1/01
Inventor 韩日畴林惠娇丘雪红颜珣
Owner GUANGDONG INST OF APPLIED BIOLOGICAL RESOURCES
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