Construction of Golden Hamster Apoa5 Gene Expression Vector and Its Expression in Different Organs

doi:10.3969/j.issn.1674-5817.2015.04.004

Abstract

Abstract: Objective To construct apolipoprotein a5 (Apoa5) expressing plamid and describe the gene expression profile of Apoa5 in golden hamster. Methods Total RNA was extracted from liver tissue of golden hamster, cDNA of Apoa5 was cloned by RT-PCR, and then subcloned into plasmid. The plasmid of pEF6/V5-His-Apoa5 was transfected into 293T cells, the expression was examined by RT-PCR and Western blot. The expression of Apoa5 in 8 organs, including brain, heart, lung, liver, spleen, kidney, testis, and ovary of golden hamster were also detected by real time PCR. Results Apoa5 expression vector was constructed. Apoa5 mRNA and APOA5 protein was successfully detected in 293T cells 48 h after transfection. Sequencing results showed that the cDNA of Apoa5 contained 1 243 bp, it encodes 366 aa. The Apoa5 gene in golden hamster has high homology with human, mouse and rat. The Apoa5 gene in golden hamster shares 75%, 84%, 84% amino acids with human, rat, and mouse. Real time PCR results showed that gene expression of Apoa5 was high in liver, brain, testis, but low in heart, lung, and kidney. Conclusion The Apoa5 expression vector was constructed successfully, and Apoa5 expression profile in golden hamster had been described. It may be as the foundation for establishing Apoa5 transgenic golden hamster and researching the function of Apoa5.

Key words: Golden hamster, Apolipoprotein A5(Apoa5), Gene expression

CLC Number:

Q95-33

DONG Wan-wei, ZHANG A-nuo, GUO Xiao-chong, WEI Zheng-li, YANG Wei, ZHAO Wen, SHEN Wei, ZHENG Zhi-hong. Construction of Golden Hamster Apoa5 Gene Expression Vector and Its Expression in Different Organs[J]. Laboratory Animal and Comparative Medicine, 2015, 35(4): 283-287.

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