CLONING AND EXPRESSION VECTOR CONSTRUCTION OF GLUTAMATE DECARBOXYLASE GENE FROM LACTOBACILLUS PLANTARUM

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ARABPOUR B.; ESMAEILI A.; RABBANI M.

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Paper Information

Journal: JOURNAL OF BABOL UNIVERSITY OF MEDICAL SCIENCES (JBUMS) Year:2016 | Volume:18 | Issue:6 Page(s): 66-72

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Title

CLONING AND EXPRESSION VECTOR CONSTRUCTION OF GLUTAMATE DECARBOXYLASE GENE FROM LACTOBACILLUS PLANTARUM

Author(s)

ARABPOUR B. | ESMAEILI A. | RABBANI M. | Issue Writer Certificate

Keywords

CLONINGQ2

GAMMA-AMINOBUTYRIC ACIDQ2

GLUTAMATE ACID DECARBOXYLASEQ1

LACTOBACILLUS PLANTARUMQ2

Abstract

BACKGROUND AND OBJECTIVE: GAMMA-AMINOBUTYRIC ACID (GABA) is a four-carbon non-protein amino acid used in the treatment of hypertension, diabetes, inflammation, and depression. GABA is synthesized by glutamic acid decarboxylase (GAD) enzyme in many organisms, including bacteria. Therefore, CLONING of this enzyme is essential to the optimization of GABA production. This study aimed to clone and construct the expression vector of GAD gene from LACTOBACILLUS PLANTARUM PTCC 1058 bacterium. METHODS: In this experimental study, we investigated the morphological, biochemical, genetic and 16s rDNA sequencing of L. plantarum PTCC 1058 strain. Genomic DNA of the bacterium was isolated and amplified using the GAD gene via polymerase chain reaction (PCR). Afterwards, the gene was inserted into the pJET1.2/blunt CLONING vector and subcloned in vector pET32a. Plasmid pET32a-gad expression vector was transformed in Escherichia coli BL21 strain, and protein expression was assessed using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). FINDINGS: Morphological, biochemical and genetic analyses of 16s rDNA sequencing indicated that the studied substrain was of the L. plantarum strain. In addition, results of nucleotide sequencing of the fragmented segment via PCR showed the presence of GAD gene. Results of colony PCR and SDS-PAGE analysis confirmed the accuracy of the CLONING and gene expression of the recombinant Escherichia coli BL21 strain.CONCLUSION: According to the results of this study, CLONING of GAD gene from L. plantarum PTCC 1058 was successful. These cloned genes could grow rapidly in prokaryotic and eukaryotic systems and be used in cost-effective culture media and even non-recyclable waste.

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APA: Copy

ARABPOUR, B., ESMAEILI, A., & RABBANI, M.. (2016). CLONING AND EXPRESSION VECTOR CONSTRUCTION OF GLUTAMATE DECARBOXYLASE GENE FROM LACTOBACILLUS PLANTARUM. JOURNAL OF BABOL UNIVERSITY OF MEDICAL SCIENCES (JBUMS), 18(6), 66-72. SID. https://sid.ir/paper/73561/en

Vancouver: Copy

ARABPOUR B., ESMAEILI A., RABBANI M.. CLONING AND EXPRESSION VECTOR CONSTRUCTION OF GLUTAMATE DECARBOXYLASE GENE FROM LACTOBACILLUS PLANTARUM. JOURNAL OF BABOL UNIVERSITY OF MEDICAL SCIENCES (JBUMS)[Internet]. 2016;18(6):66-72. Available from: https://sid.ir/paper/73561/en

IEEE: Copy

B. ARABPOUR, A. ESMAEILI, and M. RABBANI, “CLONING AND EXPRESSION VECTOR CONSTRUCTION OF GLUTAMATE DECARBOXYLASE GENE FROM LACTOBACILLUS PLANTARUM,” JOURNAL OF BABOL UNIVERSITY OF MEDICAL SCIENCES (JBUMS), vol. 18, no. 6, pp. 66–72, 2016, [Online]. Available: https://sid.ir/paper/73561/en

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