Expression of human proinsulin gene in pet 28A (+) vector and escherichia coli BL21 (DE3)

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'

Extended \ufffd?spectrum \xdf \ufffd?Lactamase in E.Coli, K.Pneumoniae and Enterobacter spp

Background:Resistance to antibiotics due to extended \ufffd?spectrum \ufffd?Lactamase (ESBL) which increased quickly, made treatment much more difficult. However, this matter was not enough to be concerned in our country. Objectives: To investigate the prevalence of ESBL producing among clinical isolates of E.coli, K.pneumoniae and Enterobacter spp and the classification of ESBLs gene by PCR. Subjects and method: 663 strains, including 248 E.coli, 393 K.pneumoniae, 22 Enterobacter spp, isolated from patients in Viet Tiep hospital (Hai Phong), Bach Mai and Pediatric hospital (Ha Noi). ESBLs were detected using modified double \ufffd?disc method. The classification of ESBLs producing strains was implemented by PCR. Results:the percentage of ESBL producing in E.coli, K.pneumoniae and Enterobacter spp is 20.2; 18.3 and 36.4%, respectively. The ESBLs producing strains were co \ufffd?resistant to most of the tested antibiotics. These strains were prevalent in intensive care units (sputum or respiratory fluid samples). TEM, SHV, CTX \ufffd?M, OXA were 87.7; 62.3; 24.6 and 12.3%, respectively. They were detected alone or in combination in the same strain. Conclusion: The rate of ESBLs producing strains is high. ESBLs were marker for multi \ufffd?drug resistance. TEM and SHV type ESBLs are most prevalent in the tested strains.

Some application of PCR in microbiology

In microbiology, PCR was applied very early and widely step by step to diagnose the etiology of the infection. Especially in case of the culture of microorganism was unsuccessfully implemented or is very difficult or patient used the antibiotic before admission because PCR can be implemented in the dead microorganism. PCR contributes to verify more correctly.

The biomedical laboratory center

The Biomedical Laboratory Center of Hanoi Medical University was established in January 17th 1997 and comprises 4 small labors: the functional tests; biochemical; immunology and genetic. According to its functions and tasks, the labor has human resources with the high technical and scientific levels. This resource originated from the faculties of Hanoi Medical University. This is an activity pattern which is suitable and convenient for staffs and students in the university.