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A kind of immunogenic cry1c recombinant protein, isolated nucleic acid molecule and application thereof

A technology for recombining proteins and nucleic acid molecules, which is applied in the field of genetic engineering and can solve the problems of long preparation time and high cost

Active Publication Date: 2021-05-28
JILIN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation time of existing cry1C antibodies is long and the cost is high. How to quickly and accurately detect cry1C protein and its related products is a problem we are facing today

Method used

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  • A kind of immunogenic cry1c recombinant protein, isolated nucleic acid molecule and application thereof
  • A kind of immunogenic cry1c recombinant protein, isolated nucleic acid molecule and application thereof
  • A kind of immunogenic cry1c recombinant protein, isolated nucleic acid molecule and application thereof

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preparation example Construction

[0042] On the other hand, the present invention also provides a method for preparing cry1C antibody, which includes the step of: immunizing animals with the above-mentioned cry1C recombinant protein. It is known that the amino acid sequence of the cry1C recombinant protein is shorter than the amino acid sequence of the full-length cry1C protein, but the cry1C recombinant protein can still immunoreact with the cry1C antibody, and the cry1C recombinant protein can be used to produce cry1C antibodies more specifically. And it has the advantages of shorter time, lower cost, stronger practicability and the like.

[0043] On the other hand, the present invention also provides a cry1C antibody prepared by the above preparation method.

no. 1 example

[0046] This embodiment provides a cry1C antibody and its preparation method.

[0047] 1 Construction of expression vector

[0048]An isolated nucleic acid molecule provided by the present invention, the nucleic acid molecule is an optimized artificially synthesized cry1C fragment, which encodes a cry1C recombinant protein with immunogenicity, and the nucleotide sequence of the nucleic acid molecule is as shown in SEQ ID No.2 shown. The nucleic acid molecule is used as a template for PCR amplification. In the PCR amplification process, a primer pair for amplifying the nucleic acid molecule is used. The primer pair includes an upstream primer as shown in SEQ ID No.3 and / or as shown in SEQ ID No. Downstream primers indicated in 4.

[0049] Specifically, the sequence of the upstream primer is: 5'- CATATG GTCGAAGCATTCAAGGA-3` (the underlined sequence is the NdeI restriction site);

[0050] The sequence of the downstream primer is: 5`- CTCGAG TTATTTCTGCGCGCGTT-3' (the underli...

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Abstract

The invention provides an immunogenic cry1C recombinant protein, which relates to the technical field of genetic engineering, and its amino acid sequence is shown in SEQ ID No.1. The amino acid sequence encoding the cry1C recombinant protein is shorter than the amino acid sequence of the full-length cry1C protein, and the cry1C recombinant protein can immunoreact with the cry1C antibody, and can be used to rapidly and efficiently prepare the cry1C antibody. The present invention also provides a nucleic acid encoding the above immunogenic cry1C recombinant protein, a pair of primers capable of amplifying or detecting the nucleic acid molecule, a vector and a host cell including the nucleic acid molecule, and in addition, the present invention also provides a cry1C Antibody and preparation method thereof, the preparation method can obtain cry1C antibody rapidly and efficiently.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to an immunogenic cry1C recombinant protein, an isolated nucleic acid molecule and applications thereof. Background technique [0002] Bacillus thuringiensis Berliner (Bt) can produce insecticidal crystal proteins (Insecticidal Crystal proteins, ICPs) harmful to a variety of insects. Bt insecticidal crystal protein (ICP) can specifically poison different insects, including Lepidoptera, Coleoptera, Diptera, mites and other pests, and is harmless to humans and animals. anti-insect protein. [0003] Rice is one of the important food crops in my country. Rice pests are one of the important limiting factors for high and stable rice yields. The annual losses to rice production in my country can account for 5% to 10% of the total output, and Lepidoptera pests are the most harmful pests to rice production. [0004] The Bt insecticidal protein encoded by cry1C gene can inhibit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/325C12N15/32C12N15/11C07K16/12C07K16/06
CPCC07K14/325C07K16/06C07K16/1278
Inventor 金永梅林秀峰
Owner JILIN ACAD OF AGRI SCI
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