FoxO1突变体转基因小鼠的建立

doi:10.16431/j.cnki.1671-7236.2018.03.007

Abstract

Abstract:

In order to explore the mechanism of FoxO1 in leptin signal transduction pathway,the recombinant plasmid pEF1α-myc-FoxO1 was constructed and transfected in 293Rb cells to detect its expression by Western blotting.The FoxO1 mutant transgenic mice were constructed by DNA pronucleus microinjection,and total genome was extracted from the mice tail and the genotypes were identified by PCR.The transcription and translation level of the FoxO1ΔDBD were detected by quantitative Real-time PCR and immunoprecipitation,and the phenotypic analysis of the FoxO1 mutant transgenic mice was conducted.The results showed that the recombinant plasmid pEF1α-myc-FoxO1ΔDBD was successfully constructed and expressed in 293Rb cells,and the 10 first generation mice and 10 lines were acquired.The method of genotypic identification of high content of G-C PCR product was established,and the genotypes of transgenic mice were detected successfully.And the expression of mutant FoxO1 was detected successfully on transcription and translation level.The result of phenotypic analysis indicated that the body weights of transgenic mice were significantly lower than wild type mice (P < 0.05),suggesting that the role of leptin was enhanced.In conclusion,the FoxO1 mutant transgenic mice models were successfully established,which provided a model for research the mechanism of FoxO1 in leptin signal transduction pathways.

Key words: FoxO1ΔDBD; transgenic mice; genotype identification; leptin; body weight

CLC Number:

Q785

ZHOU Guoli, LI Hao, LU Wanping, XU Liang, YANG Ran, YANG Guoqing. Establishment of the FoxO1 Mutant Transgenic Mice Model[J]. , 2018, 45(3): 612-619.

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Establishment of the FoxO1 Mutant Transgenic Mice Model

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