转PRL基因小鼠遗传性状分析

doi:10.11843/j.issn.0366-6964.2016.09.015

Abstract

Abstract:

In order to provide the foundation for breeding large transgenic animals in futher，in this study，the genetic traits of mammary gland-specific PRL transgenic mice produced by pronuclear microinjection，were analyzed from the genetic integrity，the copy number and the inserting site of exogenous gene.The transgenic mice deleted partially exogenous gene were screened by genomic DNA PCR method，and were verified by Southern blot to detect the genetic stability of exogenous gene.The copy number of exogenous gene in transgenic mice was detected by absolute real-time fluorescence quantitative PCR method.The inserting site of exogenous gene in transgenic mice was analyzed by Genome walking method.The results showed that the losing rate of exogenous gene fragment was 1.91%.The green fluorescence signal was observed under the long-wave UV lamp in the transgenic mice integrated higher copies of exogenous gene，while not observed in the transgenic mice integrated 2 copies of exogenous genes.The exogenous gene expressed normally in the 2 025-2 026 bp site of NW_001073945.1 gene and the 10 760 020-10 760 021 bp site of NW_001030574.1 gene.The result indicated that the exogenous gene could be inherited stablely in the transgenic mice produced by pronuclear microinjection，the expression level was positively correlated with the copy number of exogenous gene in certain range，and it was beneficial for the exogenous gene expression while there was no other genes nearby the inserting site.

Key words: transgenic animal, genetic traits, pronuclear microinjection, the inserting site of exogenous gene, copy number

CLC Number:

S865.13

REN Yan-ping，XIE Liang-liang，LI Hai-yan，SHI De-shun，LI Xiang-ping. The Genetic Traits Analysis of PRL Transgenic Mice[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(9): 1861-1867.

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The Genetic Traits Analysis of PRL Transgenic Mice

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