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目的 通过插入突变法 ,转录前加工、修饰人肝细胞生长因子 β基因 ,构建人肝细胞生长因子 β基因的可表达全序列 ,以使肝细胞生长因子 β基因可在细胞内单独表达 ,用于研究肝细胞生长因子 β基因在生物体内的生物学作用。 方法 从人肝细胞中提取总RNA ,反转录成为含肝细胞生长因子 β结构基因区序列的cDNA ,在此cDNA序列结构基因区前插入翻译起始密码子、SD序列、SD序列间隔等调节序列和限制性内切酶接头 ;PCR扩增 β基因 ;并对肝细胞生长因子 β基因可表达全序列测序。结果和结论 成功地构建并扩增了肝细胞生长因子 β基因可表达全序列。经序列分析 ,结果正确。
OBJECTIVE: To construct human hepatocyte growth factor beta gene expression full sequence by inserting mutation method, pre-transcriptional processing and modification of human hepatocyte growth factor beta gene so that hepatocyte growth factor beta gene can be expressed separately in cells for To study the biological role of hepatocyte growth factor beta gene in vivo. METHODS: Total RNA was extracted from human hepatocytes and reverse transcribed into cDNA containing the hepatocyte growth factor beta-structural region sequence. The cDNA sequence was inserted into translation initiation codon, SD sequence and SD sequence interval Sequence and restriction endonuclease linkers; PCR amplification of β gene; and expression of the full-length sequence of hepatocyte growth factor β gene sequencing. RESULTS AND CONCLUSIONS The full-length expression of the hepatocyte growth factor beta gene was successfully constructed and amplified. After the sequence analysis, the result is correct.