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Paper Information

Journal:   JOURNAL OF MAZANDARAN UNIVERSITY OF MEDICAL SCIENCES   OCTOBER 2011 , Volume 21 , Number 84; Page(s) 23 To 31.
 
Paper: 

ISOLATION OF THE PROMOTERLESS STREPTOMYCIN REGULATORY GENE (STRR2) FROM STREPTOMYCES GRISEUS

 
 
Author(s):  HOJATI ZOHREH, MOTOVALI BASHI MAJID, BIDKHORI GHOLAMREZA
 
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Abstract: 

Background and purpose: Polymerase chain reaction (PCR) is a rather quick and accurate method employed for gene detection and isolation. Primer designing is an important issue in this technique and plays a critical role in considering both the genome properties and cloning of the isolated genes. Streptomycin antibiotic is produced by Streptomyces griseus using str gene cluster with more than 25 genes. This gene cluster contains StrR gene encoding a specific protein regulator of this cluster. The pathway specific transcriptional activator then induces transcription of other genes in the str gene cluster. In this study, the researchers aimed at isolating promoterless StrR2 gene and then cloning it.
Materials and methods: To isolate promoterless StrR gene, a set of primers (StrR2) was designed. One pair of these primers (St Nes) detected the StrR from the genome. Moreover, Nested-PCR was then used to detect amplified StrR2 gene. Sites for BamHI and XbaI were designed in other primers (Str nP1and Str nP2). These primers not only amplified the StrR2 gene but also created restriction enzyme sites in the amplified fragments.
Results: Using PCR, the promoterless StrR2 gene was amplified and its structure was confirmed. This gene was successfully cloned, too. The correct structure of the recombinant plasmids was confirmed using different techniques such as gel electrophoresis, PCR and restriction digestion analysis.
Conclusion: Using this vector, one can subclone the promoterless StrR2 gene in the Streptomyces expression vectors containing inducible promoters.

 
Keyword(s): STREPTOMYCIN, PROMOTERLESS STRR2 GENE, STREPTOMYCES GRISEUS
 
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