Study on the Optimum Conditions For the Expression Of Human Recombinant Proinsulin In E.Coli - BL21(DE3) with pET - 28a(+) Vector
- Author:
Van Hong Tran
;
Phong Van Le
- Publication Type:Journal Article
- Keywords:
Expression;
Proinsulin;
E.Coli;
Appropriate conditions
- MeSH:
Expression;
Proinsulin;
E.Coli;
Appropriate conditions
- From:Journal of Medical Research
2008;0(1):56-61
- CountryViet Nam
- Language:Vietnamese
-
Abstract:
Introduction: There are thousands of diabetes sufferers worldwide. In addition, there is an increased trend in Vietnam due to economic development, increased population, lifespan, and changing lifestyle. Insulin is a hormone, which is a natural protein. It plays an important role in the transformation of glucose in human and animal blood. Note, while insulin has not been produced in Vietnam, the production of recombinant Proinsulin is a premise for insulin. This study is based on a successful design of pET - 28a (+) Vector with recombinant Proinsulin codified gene.\r\n', u'Objectives: To define the human proinsulin codified gene and study the optimum conditions for the expression of proinsulin. \r\n', u'Subjects and method: To discover the appropriate conditions for the expression of proinsulin, including initial cell density, cultural temperature, IPTG concentration, and expression time. The product of the expression was confirmed by Sodium dodecyl sulfate - polyacrylamide gel electrophoresis (SOS - PAGE) and reconfirmed by Western blotting with His - Tag antibody. \r\n', u'Results: Proinsulin expression was successfully proved by SOS - PAGE and Western blotting. Four appropriate conditions for the expression were confirmed as highlighted in the Conclusion. \r\n', u'Conclusion: The appropriate conditions for expression of proinsulin: cell density was 00600 0.6 - 1.0; the cultural temperature was 300C; IPTG concentration was 0.4 mM; the length of the culture time was 20 hours. \r\n', u'
