Extraction of Total RNA from C57BL /6J Mouse Adipose Tissue and Cloning of Perilipin Gene
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Laboratory Animal Center,Shanxi Mendical University,Taiyuan 030001,China

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    Abstract:

    Objective Using an improved method to extract high quality total RNA from mouse epididymal adipose tissue,to clone and analyze the full-length cDNA encoding perilipin. Methods According to the specification of reagent,the extraction procedure of total RNA from adipose tissue was modified to extract high quality total RNA from C57BL /6J mouse epididymal adipose tissue. The cDNA encoding perilipin was amplified by RT-PCR using the total RNA . The PCR product was cloned into pMD18-T easy vector and then transformed into E. coli JM109. The recombinant plasmid was identified with restriction enzyme digestion analysis and nucleotide sequencing. Results Using the improved method we successfully extracted high quality total RNA. The recombinant pMD18-T easy vector had a complete open reading frame of perilipin and shared 100% homology with the sequence of mRNA for perilipin reported in GenBank. Conclusions The improved extraction method of total RNA from fat tissue is feasible. The cDNA of perilipin is successfully cloned,which will be helpful for the further research on its biological function.

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History
  • Received:February 20,2011
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  • Online: December 05,2025
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