| 摘要: |
| 采用 Watson 方法由猪垂体总 mRNA 反转录合成了总 cDNA,合成产率约为12.6%。将总 cDNA 克隆到质粒 pUC19中后,转化大肠杆菌 JM107得到约2500个重组子克隆。用猪生长激素探针进行菌落原位杂交筛选得到2个阳性克隆。对这两个克隆进行多种酶切分析的结果表明,其中质粒 pWH101插入片段长度为830bp 左右,带有全长猪生长激素基因及部分5′端和3′端非编码区。用 Sanger 法分析了此基因5′端203个 bp 的序列,结果除确证己克隆了猪生长激素基因(cDNA)外,还查明了此基因5′端非编码区46bp 的序列。 |
| 关键词: 猪生长激素基因 酶切图谱 重组质粒 非编码区 菌落原位杂交 编码序列 分子克隆 直接合成 大肠杆菌 插入片段 |
| DOI： |
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| 基金项目: |
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| MOLECULAR CLONING OF cDNA OF PORCINE GROWTH HORMONE |
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| Abstract: |
| Total cDNA was synthesized from total mRNA of porcine pituitarygland by reverse trancription according to watson's method and the yield of it was about 12.6%.After recombinating the total cDNA in plasmid pUC19 and transforming E.coli JM107,2500 clones of recombinant were obtained.From these clones two positive clones were screened out by in situ colony hybridization with probes of PGH gene.Analysis of these two clones by restriction endonuelease showed that one of them is a full length gene of porcine growt... |
| Key words: Porcine Growth Hormone cDNA Molecular Cloning DNA Seguence |
