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Application of cvbv29-1 gene in reducing immunity of Drosophila and making model of immunocompromised Drosophila

A technology of low immunity and immunity, applied in the field of molecular biology and genetic engineering, can solve the problem of death of transgenic fruit flies and achieve the effect of enhanced screening

Active Publication Date: 2019-11-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, more toxic genes are usually easy to cause the death of transgenic fruit flies; therefore, finding suitable toxic genes is an important part of establishing immune-compromised fruit flies

Method used

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  • Application of cvbv29-1 gene in reducing immunity of Drosophila and making model of immunocompromised Drosophila
  • Application of cvbv29-1 gene in reducing immunity of Drosophila and making model of immunocompromised Drosophila
  • Application of cvbv29-1 gene in reducing immunity of Drosophila and making model of immunocompromised Drosophila

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Embodiment 1

[0029] 1. Construction of transgenic Drosophila

[0030] According to the sequences at both ends of the open reading frame (ORF, shown in SEQ ID NO.1) of the CvBV29-1 gene and the sequence of the multiple cloning site on the pUASttb vector, combined with the mode of action of homologous recombinase, specific primers were designed. The primer sequences are as follows:

[0031] F: 5'- TTCGTTAACAGATCTGCGGCCGC ATGGTGTTCAAAAAATCTGC-3'

[0032] R: 5'- TCCTCTAGAGGTACCTCTCGAG TTAAAGCGAAATTGCTTCTTC-3';

[0033] Trizol (Invitrogen, Carlsbad, CA, USA) was used to extract total RNA from hemocytes of Plutella xylostella after parasitism, and a cDNA library was constructed using the kit SMART RACE cDNA Amplification kit (Clontech, USA). Then, use specific primers F and R to amplify the CvBV29-1 gene fragment with restriction sites from the cDNA library constructed by PCR, and use homologous recombinase (ClonExpress II One Step Cloning Kit, Novozyme, Nanjing ) to recombine the fragment...

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Abstract

The invention discloses application of CvBv29-1 gene to reduction of fruit fly immunity and preparation of fruit fly model with low immunity. The CvBv29-1 gene is the first gene in the 29th ring of the 35th genome segment in polydnavirus of a cotesia vestalis and a base sequence of the CvBv gene is shown as SEQ ID NO. 1. By utilizing a transgenic technology, the invention transfers the CvBv29-1 gene in polydnavirus of the cotesia vestalis into a fruit fly genome and obtains transgenic fruit fly strains with stable inheritance and homozygote. After the transgenic fruit fly strains express the polydnavirus gene CvBv29-1, the phenomenon of blood cell apoptosis exists. In addition, when being infected by pathogen, the transgenic fruit fly strains has an obvious character of low immunity, and has important meaning in the aspects of research of relevant mechanism of insect immunity and enhancement of human immunological medicaments screening and the like.

Description

technical field [0001] The invention relates to the technical fields of molecular biology and genetic engineering, in particular to the application of CvBV29-1 gene in reducing the immunity of Drosophila and preparing an immune-deficient Drosophila model. Background technique [0002] Insects are currently the most prosperous species group on the earth's land, and they are an inexhaustible treasure house of resources for human beings. In recent years, the immunity of insects has received great attention in its basic and applied research. Experiments are used to study the mechanisms and signals related to insect immunity. etc. have very important practical significance. [0003] Fruit flies belong to the order Diptera, the genus Drosophila, and the family Drosophila, with about 1,000 species. Among them, Drosophila has the characteristics of short life cycle, easy breeding, fast reproduction, few chromosomes, many mutant types, and easy observation. It is widely used as an ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/34C07K14/01C12N15/85A01K67/033
CPCA01K67/0339A01K2227/706A01K2267/0387C07K14/005C12N15/8509
Inventor 黄健华王泽华陈学新时敏周思聪陈佳妮
Owner ZHEJIANG UNIV