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Paper Information

Journal:   BIOLOGICAL JOURNAL OF MICROORGANISM   WINTER 2016 , Volume 4 , Number 16; Page(s) 11 To 20.
 
Paper: 

GENE SEQUENCING, CLONING, AND EXPRESSION OF THE RECOMBINANT L- ASPARAGINASE OF PSEUDOMONAS AERUGINOSA SN4 STRAIN IN ESCHERICHIA COLI

 
 
Author(s):  BADOEI DALFARD ARASTOO*, KARAMI ZAHRA, RAMEZANI POUR NARJES
 
* SHAHID BAHONAR UNIVERSITY OF KERMAN, IRAN
 
Abstract: 

Introduction: L- asparaginase is in an excessive demand in medical applications and in food treating industries, the request for this therapeutic enzyme is growing several folds every year.
Materials and methods: In this study, a L- asparaginase gene from Pseudomonas aeruginosa strain SN4 was sequenced and cloned in E. coli. Primers were designed based on L- asparaginase from P. aeruginosa DSM 50071, which show high similarity to SN4 strain, according to 16S rRNA sequence. The L- asparaginase gene was exposed to restriction digestion with NdeI and XhoI enzymes and then ligated into pET21a plasmid. The ligated sample was transformed into competent E. coli (DE3) pLysS DH5a cells, according to CaCl2 method. The transformed E. coli cells were grown into LB agar plate containing 100
mg/ml ampicillin, IPTG (1 mM).
Results: Recombinant L- asparaginase from E. coli BL21 induced after 9 h of incubation and showed high L- asparaginase activity about 93.4 IU/ml. Recombinant L- asparaginase sequencing and alignments showed that the presumed amino acid sequence composed of 350 amino acid residues showed high similarity with P. aeruginosa L- asparaginases about 99%. The results also indicated that SN4 L- asparaginase has the catalytic residues and conserve region similar to other L- asparaginases.
Discussion and conclusion: This is the first report on cloning and expression of P. aeruginosa L- asparaginases in Escherichia coli. These results indicated a potent source of L- asparaginase for in vitro and in vivio anticancer consideration.

 
Keyword(s): L- ASPARAGINASE, CLONING, PSEUDOMONAS AERUGINOSA, SEQUENCE ALIGNMENTS
 
 
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APA: Copy

BADOEI DALFARD, A., & KARAMI, Z., & RAMEZANI POUR, N. (2016). GENE SEQUENCING, CLONING, AND EXPRESSION OF THE RECOMBINANT L- ASPARAGINASE OF PSEUDOMONAS AERUGINOSA SN4 STRAIN IN ESCHERICHIA COLI. BIOLOGICAL JOURNAL OF MICROORGANISM, 4(16), 11-20. https://www.sid.ir/en/journal/ViewPaper.aspx?id=509321



Vancouver: Copy

BADOEI DALFARD ARASTOO, KARAMI ZAHRA, RAMEZANI POUR NARJES. GENE SEQUENCING, CLONING, AND EXPRESSION OF THE RECOMBINANT L- ASPARAGINASE OF PSEUDOMONAS AERUGINOSA SN4 STRAIN IN ESCHERICHIA COLI. BIOLOGICAL JOURNAL OF MICROORGANISM. 2016 [cited 2021May16];4(16):11-20. Available from: https://www.sid.ir/en/journal/ViewPaper.aspx?id=509321



IEEE: Copy

BADOEI DALFARD, A., KARAMI, Z., RAMEZANI POUR, N., 2016. GENE SEQUENCING, CLONING, AND EXPRESSION OF THE RECOMBINANT L- ASPARAGINASE OF PSEUDOMONAS AERUGINOSA SN4 STRAIN IN ESCHERICHIA COLI. BIOLOGICAL JOURNAL OF MICROORGANISM, [online] 4(16), pp.11-20. Available: https://www.sid.ir/en/journal/ViewPaper.aspx?id=509321.



 
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