1.Sequencing of Flic genes of Salmonella typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, and S. typhimurium and application for PCR to differentiate them
Journal of Preventive Medicine 2005;15(5):36-41
All Flic genes of Salmonella typhi, S. paratyphi A, S. paratyphi B, S. paratyphi C, and S. typhimurium code for phase 1 of H antigen (H:1) (d, a, b, c and i antigen respectively). The genes were sequenced on Sanger's principle by automatic sequenser (ABI, 3100 Avant, Genetic Analyser). The primer pairs for the Salmonella species were designed on the basis of the collected sequences. The results showed that PCR with these primers can clearly differentiate the five Salmonella strains, especially between S. paratyphi B and S. typhimurium.
Polymerase Chain Reaction
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Salmonella
2.Expression of human proinsulin gene in pet 28A (+) vector and escherichia coli BL21 (DE3)
Tue Trong Nguyen ; Phung Van Le ; Chom Kyu Chong ; Lee Sang Oh
Journal of Medical Research 2007;47(2):11-15
Background: Insulin is a hormone produced by the beta \r\n', u'cells of the pancreas that permits glucose to enter cells and \r\n', u'helps the body use glucose for energy. Insulin controls the \r\n', u'amount of glucose in the blood. Insulin is produced by recombinant protein technology. Expression of human proinsulin is the first step to express insulin. Objectives:To express successfully human proinsulin gene in pET 28a vector and E.coli BL21 (DE3). Subjects and method: Human proinsulin gene was applied from human pencreas cDNA by PCR using specific PINS primer pairs which contained sites for BamH I, Xho I. Proinsulin gene was cloned into pET 28a (+) vector to form recombinant vector pET 28a-PINS then transformed into E.coli BL21 (DE3) host strain to make pET 28a-PINS/ BL21 (DE3) clone. The clone was cultured and induced by IPTG (1mM). Recombine protein was analysed by SDS-PAGE. Results: Expression vector pET 28a-PINS was constructed successfully. Proinsulin protein expressed in E.coli BL21 (DE3) was purified by ProPond-Resin (Amersham). Conclusion: Human proinsulin was produced successfully using pET 28a-PINS/ BL21 (DE3) system.\r\n', u'
Proinsulin/ pharmacology
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pharmacokinetics
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Escherichia coli