Engineering systhesized gene cry LC of pests-killing crytal protein of Bacillus thuringiensis Berliner

Abstract

The present invention is characterized by that it utilizes a series method to design and synthesize a bacillus thuringiensis Berlinear insecticidal protein gene CrylC. As compared with sequence of 1890 nucleotides of 5' end of original CrylCa5 gene the amino acid composition of said gene coded protein is not changed, but the richly-contained AT sequence, existed invented repeats and undefined eukaryotic gene intron sequence in the original gene are eliminated, C+G content is 44.64%, and the homology with original sequence is 84.0%, on the 5' end of coding sequence the guide sequence (SEQ ID No:3) is added and on the 3' end the tailing identification signal sequence (SEQ ID No.4) is added.

Description

Technical field: [0001] The invention belongs to the technical field of genetic engineering, and belongs to an innovation to the insecticidal crystal protein (ICP) gene of Bacillus thuringiensis (Bt). Specifically related to: partially changing the codon composition of the original gene, retaining the DNA sequence encoding the N-terminal toxic region of the insecticidal crystal protein in the original gene, partially removing the DNA sequence encoding the C-terminus in the original gene, and adding at the 5'end to increase gene expression The efficient guide sequence and the addition of a tail recognition sequence at the 3'end are modified to synthesize the insecticidal gene Cry1C for efficient expression in transgenic plants * . Background technique: [0002] Insect pests are an important factor in the loss of agricultural production. According to statistics, the direct economic loss caused by insect pests to agricultural production is as high as 13% ...

AI Technical Summary

Problems solved by technology

However, the insect resistance of these early Bt transgenic plants obtained by them is very weak, it is difficult to detect mRNA transcription, and the protein expression level is very low

There are many reasons for the low expression of Bt gene in plants: for example, 1, the wild Bt gene is rich in AT sequence, and the mRNA expressed in plants is unstable; 2, there may be eukaryotic gene inclusion in the wild Bt gene cleavage sites and transcription termination signal sequences, resulting in incomplete transcripts or abnormal processing of transcripts; 3. Microbes and plants have great differences in the frequency of codon usage in translation, which reduces translation efficiency; 4. True The 5'-UTR sequence of nuclear genes is very different from that of prokaryotic genes, and the 3' end of eukaryotic genes needs to be tailed to identify the signal sequence

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