[Construction of high efficient eukaryotic expression recombinant on human hepatocyte DNA synthetic stimulated factor].

Shi-gang Tang, Xian-shi Su
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引用次数: 0

Abstract

Objective: To construct the eukaryotic expression recombining vector on human The pGEM-hepatocyte DNA synthetic stimulated factor (hHDSSF) with gene cloning.

Methods: hHDSSF, a mid-recombining vector, was constructed by T-A cloning. After restriction endonuclease Not I digestion, the target fragment was subcloned into eukaryotic vector pcDNA3. 1hisB to construct the eukaryotic expression recombinant pcDNA3. 1hisB-HDSSF.

Results: The forward insert recombinant pcDNA3. 1hisB-HDSSF was screened and obtained with restriction endonuclease Kpn I digestion and it was detected by DNA sequence analysis.

Conclusion: The eukaryotic expression recombinant pcDNA3. 1hisB-hHDSSF on hHDSSF is constructed successfully, which lays a foundation for building a stable eukaryotic expression cell strain and expressing hHDSSF.

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[构建高效真核表达重组人肝细胞DNA合成刺激因子]。
目的:利用基因克隆技术构建人pgem -肝细胞DNA合成刺激因子(hHDSSF)真核表达重组载体。方法:采用T-A克隆法构建中间重组载体hHDSSF。经限制性内切酶Not I酶切后,目标片段被亚克隆到真核载体pcDNA3中。1hisB构建真核表达重组pcDNA3。1 hisb-hdssf。结果:重组pcDNA3正向插入。1hisB-HDSSF通过限制性内切酶Kpn I酶切得到,DNA序列分析检测。结论:重组pcDNA3的真核表达。成功构建了hHDSSF上的1hisB-hHDSSF,为构建稳定的真核表达细胞株,表达hHDSSF奠定了基础。
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