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:: Volume 6, Issue 2 (3-2018) ::
2018, 6(2): 309-319 Back to browse issues page
Construction of merB recombinant vector in order to bioremediation of organic mercury
Alireza Tarinejad * , Hamideh Baghi , Maryam Farshbaf Benam , Mohammad Pazhang
Department of Agricultural Biotechnology, Azarbaijan Shahid Madani University, Tabriz, Iran. , atarinejad@yahoo.com
Abstract:   (5280 Views)

Organic mercury is a sustainable form of mercury that enters the food chain of organisms and cause dangerous environmental hazards for natural ecosystem. Mercury stability and high cost conventional refining methods are disturbing factors. In this field, biological methods such as the use of bacteria or enzyme based for microbial remediation provided low-cost strategy and environmental lover of bioremediation. In this research, merB gene isolated from resistance bacteria by PCR technique by specific primers containing NcoI and NdeI restriction sites, so after amplification, it cut with restriction enzymes and after purification of gene, ligation was performed into PET28 a+ expression vector. PET28a+-merB recombinant vector was transferred into E. coli strain TOP10 by heat shock. The recombinant bacteria were selected in kanamycin selective medium. The presence of gene in transformed bacteria was confirmed by PCR on plasmid and by enzymatic digestion. Results confirmed the accuracy of cloning of merB gene into this expression vector. In order to the expresse its protein, the recombinant vector was transferred into E. coli BL21 (DE3). Expression of the protein was confirmed by SDS-PAGE method. Finally, the growth of E.coli strain BL21 containing the recombinant vector with E coli BL21 bacteria including empty vector was measured by adding methyl mercury into the environment during 48 hours. Growth ability of E.coli containing the empty vector indicates lack of bacteria resistance to mercury, on the other hand; merB protein in transformed bacteria increased their resistance to methyl mercury in the environment. Therefore, bioremediation of organic mercury from pollutant environments could be possible by using this methodology.
 

Keywords: Organic mercury, reducing gene of organic mercury, recombinant vector
Full-Text [PDF 1161 kb]   (1996 Downloads)    
Type of Study: Research | Subject: Biosafety
Received: 2016/12/22 | Accepted: 2018/03/10 | Published: 2018/03/14
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Tarinejad A, Baghi H, Farshbaf Benam M, Pazhang M. Construction of merB recombinant vector in order to bioremediation of organic mercury. Genetic Engineering and Biosafety Journal 2018; 6 (2) :309-319
URL: http://gebsj.ir/article-1-158-en.html


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Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 6, Issue 2 (3-2018) Back to browse issues page
دوفصل نامه علمی-پژوهشی مهندسی ژنتیک و ایمنی زیستی Genetic Engineering and Biosafety Journal
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