CLONING & EXPRESSION OF GENE FOR THE VITAMIN B12 RECEPTOR IN E. COLI O157:H7

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GHAJAR KOUHESTANI M.; ALIPOUR M.; REZAE A.

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Journal: Biotechnological Journal of Environmental Microorganisms Year:2012 | Volume: | Issue: Page(s): 55-62

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Information Journal Paper

Title

CLONING & EXPRESSION OF GENE FOR THE VITAMIN B12 RECEPTOR IN E. COLI O157:H7

Author(s)

GHAJAR KOUHESTANI M. | ALIPOUR M. | REZAE A. | Issue Writer Certificate

Keywords

VITAMIN B12Q2

BTUBQ2

E. COLI O157 H7Q2

Abstract

The BTUB gene of E. coli O157:H7 is made up of 1845 base pairs. This gene codes BTUB protein that is called VITAMIN B12 transporter. BTUB protein is necessary for absorbing VITAMIN B12 in E. coli. This transporter has two domains: The outer membrane b barrel domain and plug domain. The plug domain blocks b barrel channel. VITAMIN B12 in presence of calcium is absorbed to BTUB with high affinity. Extracellular loops L2 and L3 bind to VITAMIN B12. The gene coding for BTUB protein from E. coli O157:H7 was amplified. This gene was cloned and expressed as C-terminal His (6) -tagged protein. BTUB gene by designing special primer reproduced with PCR. PCR product and selected vector cutting with limited enzyms and connected together and produce construct which transformed to E. coli BL21 (DE3). Then, transformed cells after induction with IPTG detect just one band with molecular weight about 66 KDalton which indicates BTUB protein molecular weight. The SDS-PAGE analysis of the total protein revealed only 1 distinct bands, with molecular masses of 66 kDalton. purification of BTUB helped reveal its appropriate molecular mass. Regarding to this issue that protein has many Extracellular lopes, therefore, One of the purpose of this study was producing and recognizing periplasmic connecting protein of (BtuB) B12 vitamin transferred of E. coli O157:H7 and the other purpose is determination of proper molecular weight of BTUB protein.

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APA: Copy

GHAJAR KOUHESTANI, M., ALIPOUR, M., & REZAE, A.. (2012). CLONING & EXPRESSION OF GENE FOR THE VITAMIN B12 RECEPTOR IN E. COLI O157:H7. BIOTECHNOLOGICAL JOURNAL OF ENVIRONMENTAL MICROORGANISMS, 6(1), 55-62. SID. https://sid.ir/paper/242204/en

Vancouver: Copy

GHAJAR KOUHESTANI M., ALIPOUR M., REZAE A.. CLONING & EXPRESSION OF GENE FOR THE VITAMIN B12 RECEPTOR IN E. COLI O157:H7. BIOTECHNOLOGICAL JOURNAL OF ENVIRONMENTAL MICROORGANISMS[Internet]. 2012;6(1):55-62. Available from: https://sid.ir/paper/242204/en

IEEE: Copy

M. GHAJAR KOUHESTANI, M. ALIPOUR, and A. REZAE, “CLONING & EXPRESSION OF GENE FOR THE VITAMIN B12 RECEPTOR IN E. COLI O157:H7,” BIOTECHNOLOGICAL JOURNAL OF ENVIRONMENTAL MICROORGANISMS, vol. 6, no. 1, pp. 55–62, 2012, [Online]. Available: https://sid.ir/paper/242204/en

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