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OALib Journal期刊
ISSN: 2333-9721
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人血小板衍生生长因子b链基因酵母表达载体的构建
, PP. 273-276
Keywords: 血小板源生长因子,遗传载体/化学合成,rt-pcr,酵母
Abstract:
目的构建带信号肽和不带信号肽的人血小板衍生生长因子b(pdgf-b)链基因酵母表达载体,探讨该基因在酵母中的表达效率和活性。方法利用逆转录-聚合酶链反应(rt-pcr)从人血管内皮细胞总rna中扩增出pdgf-b的cdna,克隆入pgem-t载体,进行序列测定。进而克隆至酵母表达质粒pmetb和pmetαa并进行重组子双酶切鉴定。结果扩增获得了含信号肽(578bp)和不含信号肽(340bp)的编码pdgf-b链基因;构建了pdgf-b的酵母表达载体pmetb-pdgfb1,pmetαa-pdgfb2。测序结果显示,重组载体中目的基因序列与genebank公布的序列一致。结论人血小板衍生生长因子b链基因酵母表达载体构建成功,为进一步在酵母中的高效表达奠定基础。
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