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Vector for controlling gender of animals and application thereof

A vector and gender technology, applied in the field of siRNA recombinant expression vector, can solve the problems of little difference in performance, limited promotion, and difficulty in determining

Inactive Publication Date: 2011-10-05
SHIHEZI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far, except for the difference in the DNA content of X and Y sperm, the difference in other aspects is minimal or difficult to determine
Although the accuracy of the sperm obtained by flow cytometry is high, the conception rate is low, and the instrument is expensive, which limits the promotion of this technology

Method used

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  • Vector for controlling gender of animals and application thereof
  • Vector for controlling gender of animals and application thereof
  • Vector for controlling gender of animals and application thereof

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Embodiment Construction

[0022] The present invention will be described in detail below in conjunction with specific embodiments.

[0023] 1. Zfy gene RNAi recombinant vector construction

[0024] The RNAi target sequence of the Zfy gene was designed, and two siRNAs were screened out (see Table 1). The cleavage sites of restriction endonucleases BamHI and HindIII were introduced at the 5' and 3' ends respectively, and hairpin RNA (Short Hairpin RNA, shRNA) oligonucleotide chains and their complementary chains were synthesized (see Table 2).

[0025] The upstream and downstream chains of Zfy205 (control group) and Zfy2012 (experimental group) were combined respectively, and then connected into the pSilencer5.1-H1Retro vector (Ambion Shanghai Jitai Bioengineering Co., Ltd.) double-digested with BamHI and HindIII (TaKaRa company) , constructed into siRNA expression vectors pSilencer5.1 / Zfy 205 (p205 for short), pSilencer5.1 / Zfy2012 (p2012 for short), and transformed into E.coli Top10 competent cells (Be...

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Abstract

The invention discloses a vector for controlling gender of animals and a preparation method thereof. The sequences of the vector provided by the invention contain one or two selected from the following segments of: SEQ 1:5'-AGCTTCAAATTCAAGAGATTTGAAGCTCACACTGTTGTTTTTTGGAAA-3' SEQ 2:5'-CAACAGTGTGAGCTTCAAATCTCTTGAATTTGAAGCTCACACTGTTGG-3'.

Description

technical field [0001] The invention relates to a carrier for controlling animal sex, in particular to a Zfy gene siRNA recombinant expression carrier and application thereof. Background technique [0002] Over the years, people have done a lot of research on the differences between X and Y sperm, and tried to use these differences to separate the sperm and use the resulting sperm to fertilize, so that the sex of the embryo is determined at the time of fertilization. But so far, except that the DNA content of X and Y sperm has a certain difference, the difference in other aspects is little or difficult to determine. Although the sperm obtained by flow cytometry has high accuracy, the conception rate is low, and the instrument is expensive, which limits the promotion of this technology. However, if the key specific mRNAs of X and Y sperm can be found, combined with the contemporary emerging RNAi technology, it is expected to achieve sex control of animals easily and quickly ...

Claims

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Application Information

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IPC IPC(8): C12N15/85A61K48/00A61P15/00
Inventor 贾斌彭强李仁燕班谦惠文巧赵宗胜申红王开胜刘小军
Owner SHIHEZI UNIVERSITY