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含人表皮生长因子（hEGF）的酵母重组表达载体的构建

Construction of Yeast Recombinant Expression Vector Containing Human Epidermal Growth Factor (hEGF).

作者信息

Mohammadian Jamal, Mansoori-Derakhshan Sima, Mohammadian Masood, Shekari-Khaniani Mahmoud

机构信息

Department of Clinical Biochemistry, Division of Medical Biotechnology, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.

出版信息

Adv Pharm Bull. 2013;3(2):473-6. doi: 10.5681/apb.2013.079. Epub 2013 Aug 20.

DOI:10.5681/apb.2013.079
PMID:24312882
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC3848235/
Abstract

PURPOSE

The objective of this study was construction of recombinant hEGF-pPIC9 which may be used for expression of recombinant hEGF in following studies.

METHODS

EGF cDNA was purchased from Genecopoeia Company and used for PCR amplification. Prior to ligation, the PCR product and pPIC9 vector was digested with EcoRI and XhoI and ligated in pPIC9 vector and subjected to colony PCR screening and sequencing analysis.

RESULTS

PCR amplification of EGF cDNA using recombinant hEGF-pPIC9 vector as template was concluded in amplification of 197bp fragment. Construction of recombinant hEGF-pPIC9 of EGf gene was verified by PCR and sequencing.

CONCLUSION

Construction of Recombinant hEGF-pPIC9 was the primary stage for production and expression of EFG in the future study.

摘要

目的

本研究的目的是构建重组hEGF-pPIC9，其可用于后续研究中重组hEGF的表达。

方法

从Genecopoeia公司购买EGF cDNA并用于PCR扩增。在连接之前，将PCR产物和pPIC9载体用EcoRI和XhoI进行酶切，并连接到pPIC9载体中，然后进行菌落PCR筛选和测序分析。

结果

以重组hEGF-pPIC9载体为模板进行EGF cDNA的PCR扩增，得到了197bp的片段。通过PCR和测序验证了Egf基因重组hEGF-pPIC9的构建。

结论

重组hEGF-pPIC9的构建是未来研究中EFG生产和表达的初级阶段。

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