猪生长激素促分泌素受体基因真核表达载体的构建及鉴定

Abstract

Abstract: This test was intended to construct eukaryotic expression vector of porcine growth hormone secretagogue receptor (pGHS-R), and observe its expression in transiently transfected HEK293T cells. The coding DNA sequence of GSH-R gene was amplified by SOE-PCR technology from porcine genomic DNA and cloned into pcDNA3.1(+) vector, then inserted myc-tag. The positive clone was confirmed by enzyme digestion and sequencing. The expression vector was transiently transfected into HEK293T cells. The expression of pGHS-R protein was identified by Western blotting. The results showed that the CDS of pGHS-R was successfully cloned; the construction of pcDNA3.1(+)-myc/pGHS-R was correct and its expression of pGHS-R protein was confirmed by Western blotting. The results showed that the eukaryotic expression vector pcDNA3.1(+)-myc/pGHS-R was successfully constructed and pGHS-R protein was successfully expressed. The study laid a foundation for further study on GHS-R gene function in vitro.

Key words: growth hormone secretagogue receptor; eukaryotic expression; splicing overlapping extension PCR; human emborynic kidney

SHI Lin, LIN Shuai, ZHOU Dong-qing, SUN Yan-jun, WANG Zhi-qiang. Construction and Identification of Eukaryotic Expression Vector of Porcine Growth Hormone Secretagogue Receptor Gene[J]. .

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Construction and Identification of Eukaryotic Expression Vector of Porcine Growth Hormone Secretagogue Receptor Gene

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