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

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BADOEI DALFARD ARASTOO; KARAMI ZAHRA; RAMEZANI POUR NARJES

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Journal: Journal of Microbial Biology Year:2016 | Volume:4 | Issue:16 Page(s): 11-20

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Title

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 | Issue Writer Certificate

Keywords

L- ASPARAGINASEQ2

CLONINGQ2

PSEUDOMONAS AERUGINOSAQ2

SEQUENCE ALIGNMENTSQ2

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.

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APA: Copy

BADOEI DALFARD, ARASTOO, KARAMI, ZAHRA, & RAMEZANI POUR, NARJES. (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. SID. https://sid.ir/paper/237469/en

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[Internet]. 2016;4(16):11-20. Available from: https://sid.ir/paper/237469/en

IEEE: Copy

ARASTOO BADOEI DALFARD, ZAHRA KARAMI, and NARJES RAMEZANI POUR, “GENE SEQUENCING, CLONING, AND EXPRESSION OF THE RECOMBINANT L- ASPARAGINASE OF PSEUDOMONAS AERUGINOSA SN4 STRAIN IN ESCHERICHIA COLI,” BIOLOGICAL JOURNAL OF MICROORGANISM, vol. 4, no. 16, pp. 11–20, 2016, [Online]. Available: https://sid.ir/paper/237469/en

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