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Method for producing transgenic pigs through overexpression HOXA10 genes

A HOXA10, transgenic pig technology, applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc., can solve the problems of slow progress, little effect, low heritability, etc., to achieve simple operation, low cost, positive The effect of short detection time

Inactive Publication Date: 2015-04-22
HUAZHONG AGRI UNIV
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Problems solved by technology

However, since the litter size trait is affected by various factors such as fertilization rate, ovulation rate, and embryo implantation, it is also a low heritability trait, with a heritability of only 0.10-0.15. Improvement through traditional breeding methods is slow, With little success

Method used

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  • Method for producing transgenic pigs through overexpression HOXA10 genes
  • Method for producing transgenic pigs through overexpression HOXA10 genes
  • Method for producing transgenic pigs through overexpression HOXA10 genes

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Experimental program
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Embodiment 1

[0039] Embodiment 1 utilizes the method for overexpressing HOXA10 gene to prepare transgenic pig

[0040] The reagents used in this test include: the reagents used in the cell test were purchased from Sigma Company unless otherwise specified, the plasmid extraction kit and genomic DNA extraction kit were purchased from Tiangen Biochemical Technology (Beijing) Co., Ltd. unless otherwise specified, all restrictions Endonucleases were purchased from NEB unless otherwise specified.

[0041] Step 1. Enzyme digestion verification and linearization of pc3.1-HOXA10-DNA vector

[0042] The plasmid pc3.1-HOXA10-DNA (see figure 1 ) and measure the concentration. The vector plasmid is a plasmid constructed in our laboratory earlier: the HOXA10 gene promoter fragment is inserted between the MluI restriction site at 229bp of the vector pcDNA3.1plus and the HindIII restriction site at 912bp, see sequence table SEQ ID NO: 1 The promoter sequence at 234-1301bp, or see Figure 7The sequenc...

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Abstract

The invention belongs to the technical field of pig transgenosis and particularly relates to a method for producing transgenic pigs through overexpression HOXA10 genes. The method is characterized in that constructed HOXA10 gene expression carriers are subjected to lipofection and sulfate (G418) screening to obtain HOXA10 transgenic cell lines with stable expression; the HOXA10 transgenic cell lines are transplanted into denucleated pig oocytes with HOXA10 transgenic pig fibroblasts as nuclear donors through an artificial nuclear transplantation method to form reconstructed embryos; the reconstructed embryos are transplanted into a receptor sow oviduct, and after a sow gives birth to the HOXA10 transgenic pigs and molecular identification is conducted, masculine HOXA10 transgenic pigs are screened out. The transgenic pigs obtained through the method provide materials for research about the mechanism of improving the reproductive capacity through the HOXA10 genes and provide an animal model for revealing other functions of the HOXA10 genes.

Description

technical field [0001] The invention belongs to the technical field of pig transgenics, and in particular relates to a method for preparing transgenic pigs by overexpressing the HOXA10 gene. Background technique [0002] Pig farming is an important part of the animal husbandry production system and occupies a pivotal position in the animal husbandry industry. For pig producers, pig reproductive traits, especially litter size, are one of the important factors affecting production efficiency. However, since the litter size trait is affected by various factors such as fertilization rate, ovulation rate, and embryo implantation, it is also a low heritability trait, with a heritability of only 0.10-0.15. Improvement through traditional breeding methods is slow, With little success. Although genetic improvement of litter size is difficult, it is particularly important to develop and utilize new genetic improvement technologies because of its high economic benefits. [0003] Sin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/873C12N15/85A01K67/027
Inventor 赵书红肖倩赵长志林瑞意经璐李新云李长春余梅成宏
Owner HUAZHONG AGRI UNIV
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