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Pig Xist (X inactive specific transcript) gene sequence, pig anti-Xist siRNA (small interfering ribose nucleic acid) sequence and method for improving pig cloning efficiency through inhibiting Xist gene expression

A gene expression and gene sequence technology, applied in the field of genetic engineering, can solve the problem of unpublished Xist gene sequence, and achieve the effect of improving cloning efficiency and good repeatability

Active Publication Date: 2013-06-26
SOUTH CHINA AGRI UNIV
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Problems solved by technology

However, no similar experiment has been reported on pigs, and the sequence of the pig's Xist gene has not been published.

Method used

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  • Pig Xist (X inactive specific transcript) gene sequence, pig anti-Xist siRNA (small interfering ribose nucleic acid) sequence and method for improving pig cloning efficiency through inhibiting Xist gene expression
  • Pig Xist (X inactive specific transcript) gene sequence, pig anti-Xist siRNA (small interfering ribose nucleic acid) sequence and method for improving pig cloning efficiency through inhibiting Xist gene expression
  • Pig Xist (X inactive specific transcript) gene sequence, pig anti-Xist siRNA (small interfering ribose nucleic acid) sequence and method for improving pig cloning efficiency through inhibiting Xist gene expression

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Embodiment Construction

[0058] The present invention will be further described below in conjunction with specific examples, but the specific examples do not limit the present invention in any way.

[0059] 1. Electronic cloning of pig Xist gene

[0060] Using the MegAlign program of DNAStar (www.dnastar.com / ), Bostaurus (bovine, NCBI serial number: NR 001464.2), Homo sapiens (human, NCBI serial number: NG 016172.1), Mus musculus (mouse, NCBI sequence number: AJ421479.1) for homology analysis of the nucleotide sequence of Xist, a relatively conservative sequence was found. Based on the relatively conserved sequence found on the UCSC website, BLAST analysis was performed with the pig Nov.2009 (SGSC Sscrofa9.2 / susScr2) database, and this sequence was located on the pig X chromosome, and extended forward and backward based on this , to obtain a sequence of about 40kb. The 40kb sequence obtained above was analyzed by BLAST with the XIST / Xist sequences of bovine, human and mouse respectively, and a 30kb ...

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Abstract

The invention discloses a complete pig Xist (X inactive specific transcript) gene sequence and a method for improving the pig cloning efficiency. The pig Xist gene sequence is obtained through electronically cloning the pig Xist gene sequence by utilizing a biological informatics method and checking and analyzing the sequence. The method comprises the following steps of: designing and synthesizing anti-Xist siRNA (small interfering ribose nucleic acid) sequences, screening the effective anti-Xist siRNA sequence by utilizing a molecular biological technology, utilizing anti-Xist siRNA to inhibit nuclear transfer reconstructed embryos or Xist gene expression of nuclear transfer somatic cell donors so as to improve the development quality of pig cloning embryos, and thus realizing a purpose of improving the pig cloning efficiency. According to the method, the complete pig Xist sequence and the anti-Xist siRNA sequence are obtained at first; and moreover, by utilizing the method, the ectogenic quality of the pig cloning embryos can be improved, and the improved pig cloning efficiency has a significant meaning in the fields of livestock production, scientific research or medical science.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to transgenic engineering technology, in particular to a method for improving pig cloning efficiency. Background technique [0002] In 1938, the German scientist Spemanm first proposed and carried out the experiment of cell nucleus transfer. Afterwards, a large number of scientists invested in the experiment of cloning animals by nuclear transfer. It was not until 1997 that the Roslin Institute in the United Kingdom announced that the cloned sheep "Dolly" was successfully bred, indicating that somatic cells It can be used as a nucleus donor cell for nuclear transplantation, which provides a theoretical basis for the vigorous development of cloning technology and opens a new page in the history of animal cloning. Subsequently, a large number of cloned animals were born one after another, and in 2000, the world's first somatic cell nuclear transplantation pig using pig fet...

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Application Information

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IPC IPC(8): C12N15/12C12N15/113C12N15/89C12N15/85A01K67/027
Inventor 黄志华刘德武李紫聪袁玉娟吴珍芳
Owner SOUTH CHINA AGRI UNIV
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