[Home ] [Archive]   [ فارسی ]  
:: Main :: About :: Current Issue :: Archive :: Search :: Submit :: Contact ::
Main Menu
Journal Information::
Indexing Sources::
Editorial Board::
Executive Members::
Articles Archive::
Instruction to Authors::
Contact Us::
Site Facilities::
Search in website

Advanced Search
Receive site information
Enter your Email in the following box to receive the site news and information.
:: Volume 23, Issue 3 (10-2021) ::
J Gorgan Univ Med Sci 2021, 23(3): 84-90 Back to browse issues page
Cloning and Expression of Urate Oxidase Gene Isolated from Marine Streptomyces in Escherichia coli Origami Bacteria
Nayyereh Sadat Jenaban1 , Elahe Ali Asgari * 2, Kumarss Amini3
1- M.A in Biology, Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran.
2- Assistant Professor, Department of Biology, East Tehran Branch, Islamic Azad University, Tehran, Iran. , e.asgari@gmail.com
3- Associate Professor, Department of Microbiology, School of Basic Sciences, Saveh Branch, Islamic Azad University, Saveh, Iran.
Abstract:   (4238 Views)
Background and Objective: Streptomyces are gram-positive and aerobic bacterial strains that are isolated from different sources. Streptomyces have the ability to produce secondary metabolites and biologically active substances and are therefore very important in the field of biocontrol. Urate oxidase is a microbial enzyme product that can be extracted from a variety of sources, including streptomycin. In the present study, cloning of the urate oxidase gene isolated from seawater streptomycosis was performed in Escherichia coli Origami bacteria.
Methods: In this descriptive study, a total of 60 water and sediment samples were collected from different depths of the Caspian Sea coast in Mazandaran province, Iran. The Geram, staining methyl red, VP, citrate, starch hydrolysis, casein hydrolysis, nitrate reduction, oxidase and catalase tests were performed to identify and isolate Streptomyces. The urate oxidase gene was cloned using the T-A cloning method using the PTG-19 vector inside the host of Escherichia coli Origami. The expression of cloned genes in recombinant colonies was investigated by Real-Time PCR. The phylogenetic tree was drawn using clustalX and Mega5 software.
Results: Screening of marine water samples identified 12 isolated streptomyces, all of which had the urate oxidase gene. The expression of urate oxidase gene in Escherichia coli Origami was confirmed by Real-Time PCR. The results of phylogenetic studies identified some close relatives of Streptomyces as candidates for subsequent studies.
Conclusion: Streptococcus bacteria can be considered as a rich source of secondary metabolites with many applications and can be used as a native to produce the enzyme urate oxidase. By using different cloning hosts and examining optimal production conditions, this strain can be a candidate for future studies to develop antimicrobial drugs and compounds.
Keywords: Cloning [MeSH], Urate Oxidase [MeSH], Real Time PCR [MeSH], Caspian Sea [MeSH]
Article ID: Vol23-44
Full-Text [PDF 602 kb]   (11614 Downloads)    
Type of Study: Original Articles | Subject: Medical Biotecnology
1. Cheng X, Liu F, Zhang Y, Jiang Y. Cloning and expression of a urate oxidase and creatinine hydrolase fusion gene in Escherichia coli. Ren Fail. 2013; 35(2): 275-8. DOI: 10.3109/0886022X.2012.745787 [DOI] [PubMed]
2. Asghari N, Shahzadeh Fazeli SA, Amoozegar MA. [Screening and biosynthesis of Uricase enzyme-producing salt-loving arc strains]. M.A Thesis. University of Science and Culture. Faculty of Basic Sciences. 2016. [Persian]
3. Tork SE, Aly MM, Elsemin O. A new l-glutaminase from Streptomyces pratensis NRC 10: Gene identification, enzyme purification, and characterization. Int J Biol Macromol. 2018 Jul; 113: 550-57. DOI: 10.1016/j.ijbiomac.2018.02.080 [DOI] [PubMed]
4. Ebrahimi Sh, JameiR, Imani M. [Cloning and expression of Uricase gene of Baciluss subtilis in bacterial strain (BL21) E.coli]. M.A Thesis. Faculty of Sciences. Urmia University. 2014.
5. Satoh E, Niimura Y, Uchimura T, Kozaki M, Komagata K. Molecular cloning and expression of two alpha-amylase genes from Streptococcus bovis 148 in Escherichia coli. Appl Environ Microbiol. 1993 Nov; 59(11): 3669-73. DOI: 10.1128/aem.59.11.3669-3673.1993 [DOI] [PubMed]
6. Kämpfer P. The Family Streptomycetaceae, Part I: Taxonomy. In: Dworkin M, Falkow S, Rosenberg E, Schleifer KH, Stackebrandt E. (eds). The Prokaryotes. New York: Springer. 2006; pp: 538-604. DOI: 10.1007/0-387-30743-5_22 [Article] [DOI]
7. Mabrouk AM, Hamed ER, Farag MM, Ahmed NE. Purification and Characterization of Uricase Enzyme Produced by Gliomastix gueg. Gate 2 Biotech. 2010; 2(11): 1-13. [View at Publisher]
8. Song J, Lee SC, Kang JW, Baek HJ, Suh JW. Phylogenetic analysis of Streptomyces spp. isolated from potato scab lesions in Korea on the basis of 16S rRNA gene and 16S-23S rDNA internally transcribed spacer sequences. Int J Syst Evol Microbiol. 2004 Jan; 54(Pt 1): 203-209. DOI: 10.1099/ijs.0.02624-0 [DOI] [PubMed]
9. Ohe T, Watanabe Y. Purification and properties of urate oxidase from Streptomyces cyanogenus. J Biochem. 1981 Jun; 89(6): 1769-76. DOI: 10.1093/oxfordjournals.jbchem.a133376 [DOI] [PubMed]
10. Ciesarova Z, Kiss E, Boegl P. Impact of L-asparaginase on acrylamide content in potato products. J Food and Nutr Res. 2006;45(4):141-46. [Article]
11. Imani M, Pazhang M, Mirzaeinia S. [Cloning and Expression of Therapeutic Enzyme, Aspergillus Flavus Uricase in E. coli]. J Adv Med Biomed Res. 2016; 24(106): 109-121. [Article in Persian] [View at Publisher]
12. Shaaban MI, Abdelmegeed E, Ali YM. Cloning, Expression, and Purification of Recombinant Uricase Enzyme from Pseudomonas aeruginosa Ps43 Using Escherichia coli. J Microbiol Biotechnol. 2015 Jun; 25(6):887-92. DOI: 10.4014/jmb.1410.10041 [DOI] [PubMed]
Send email to the article author

XML   Persian Abstract   Print

Download citation:
BibTeX | RIS | EndNote | Medlars | ProCite | Reference Manager | RefWorks
Send citation to:

Jenaban N S, Ali Asgari E, Amini K. Cloning and Expression of Urate Oxidase Gene Isolated from Marine Streptomyces in Escherichia coli Origami Bacteria. J Gorgan Univ Med Sci 2021; 23 (3) :84-90
URL: http://goums.ac.ir/journal/article-1-3835-en.html

Rights and permissions
Creative Commons License This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
Volume 23, Issue 3 (10-2021) Back to browse issues page
مجله دانشگاه علوم پزشکی گرگان Journal of Gorgan University of Medical Sciences
Persian site map - English site map - Created in 0.05 seconds with 36 queries by YEKTAWEB 4652