CLONING, SEQUENCE ANALYSIS, AND EXPRESSION OF L-ASPARAGINASE OBTAINED FROM ERWINIA CHRYSANTHEMI

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AFRASIABI RAZIEH; AGHAIYPOUR KHOSROW; KAFILZADEH FARSHID; SAFAVIYEH SADIGHE

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Journal: JOURNAL OF MICROBIAL WORLD Year:2010 | Volume:3 | Issue:1 (6) Page(s): 7-15

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Title

CLONING, SEQUENCE ANALYSIS, AND EXPRESSION OF L-ASPARAGINASE OBTAINED FROM ERWINIA CHRYSANTHEMI

Author(s)

AFRASIABI RAZIEH | AGHAIYPOUR KHOSROW | KAFILZADEH FARSHID | SAFAVIYEH SADIGHE | Issue Writer Certificate

Keywords

L- ASPARAGINASEQ3

ERWINIA CHRYSANTHEMIQ3

PAED4Q3

SDS-PAGEQ2

Abstract

Background and objectives: L-asparaginases enzyme is an effective drugs for treatment of lymphoblasticleukemia and Non-Hodgkin's lymphoma. Escherichia coli and Erwinia strains specially ERWINIA CHRYSANTHEMI are more important producers of this enzyme. The aim of this study was survey on L-asparaginase enzyme obtained from ERWINIA CHRYSANTHEMI DSM4610.Materials and methods: After bacterial genomic DNA extraction, a PCR procedure was performed with specific primers, Echr Asn F and Echr Asn R1. At the next step, the amplified DNA segment were ligated in a pTZ57R/T vector and after transformation into E. coli DH5a, the recombinant plasmids were extracted. After sequencing, the results were analyzed by DNAMAN software. By designin its expression primers, the gene was cloned in a PAED4 vector and after transformation into E.coli BL21 (DE3) cells, its expression was assessed by SDS-PAGE analysis.Results: The sequence of the isolated gene from ERWINIA CHRYSANTHEMI DSM4610 was different from other similar genes in Gene Bank and was assigned with the accession number: JF972567 in Gene Bank. The most expression was found in the condition of 0.5 mM of IPTG, LB broth media and 37oC. SDS-PAGE analysis showed 37.5 KD protein.Conclusion: Results revealed that the L-asparaginase enzyme obtained of ERWINIA CHRYSANTHEMI has proper features for further study as an antileukemia enzyme.

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APA: Copy

AFRASIABI, RAZIEH, AGHAIYPOUR, KHOSROW, KAFILZADEH, FARSHID, & SAFAVIYEH, SADIGHE. (2010). CLONING, SEQUENCE ANALYSIS, AND EXPRESSION OF L-ASPARAGINASE OBTAINED FROM ERWINIA CHRYSANTHEMI. JOURNAL OF MICROBIAL WORLD, 3(1 (6)), 7-15. SID. https://sid.ir/paper/189280/en

Vancouver: Copy

AFRASIABI RAZIEH, AGHAIYPOUR KHOSROW, KAFILZADEH FARSHID, SAFAVIYEH SADIGHE. CLONING, SEQUENCE ANALYSIS, AND EXPRESSION OF L-ASPARAGINASE OBTAINED FROM ERWINIA CHRYSANTHEMI. JOURNAL OF MICROBIAL WORLD[Internet]. 2010;3(1 (6)):7-15. Available from: https://sid.ir/paper/189280/en

IEEE: Copy

RAZIEH AFRASIABI, KHOSROW AGHAIYPOUR, FARSHID KAFILZADEH, and SADIGHE SAFAVIYEH, “CLONING, SEQUENCE ANALYSIS, AND EXPRESSION OF L-ASPARAGINASE OBTAINED FROM ERWINIA CHRYSANTHEMI,” JOURNAL OF MICROBIAL WORLD, vol. 3, no. 1 (6), pp. 7–15, 2010, [Online]. Available: https://sid.ir/paper/189280/en

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