含第一内含子猪生长激素cDNA基因在COS7细胞中的表达

Construction of Expression Vector of pGH cDNA with First Intron and its Expression in COS7 Cells

以CMV真核生物启动子构建了含第一内含子猪生长激素(pGH)cDNA基因(pGHcDNA in)的表达载体,经DEAE 葡聚糖介导在COS7细胞进行了瞬时表达,通过PP95生物学测定法测定,证明含第一内含子pGHcDNA基因能够在真核细胞中进行分泌性表达,表达出的pGH具有生物学活性。这说明通过该基因转录出的前体mR NA能够在COS7细胞中进行正确剪接,从而翻译出具有生物活性的pGH。加入第一内含子在一定程度上可提高pGHcDNA基因的表达效率。

It has been reported that the intron of eucaryotic genes have the regulatory elements for gene expressions.The porcine growth hormone cDNA with first intron was constructed in our lab.In order to learn the effect of the first intron on gene expression,the expression vector of pGHcDNA with first intron(pGH cDNAin)under the control of CMV prompter was constructed.By transient expression mediated with DEAEDextran in COS7 cells and detection with PP95 biological method,the pGH cDNAin could be secretary expressed in eucaryotic cells,and expressed pGH showed biological activity.This result indicated that primary mRNA transcribed from pGH cDNAin gene could be spliced correctly in COS7 cells,therefore,was translated into pGH with biological activity.The expression level of pGH was increased in a certain extent by inserting the first intron into pGH cDNA gene.