Cloning and expression of Iranian isolate of Apple chlorotic leaf spot virus coat protein gene in Escherichia coli

Q: How can I download an article?

To download an article from SID, first log in to the site, search for the article title, and click on the 'Download Article' option.

Q: How can I download an ISI article?

To download an ISI article on SID, enter the keyword or article title in the search bar, view the relevant results, click on the desired article, and select the 'Download Article' option.

Q: How can I access the SID database?

To access the SID database, visit SID.ir, create an account, and log in to access scientific resources.

Q: Is downloading articles from SID free?

Some articles on SID are available for free, while others require payment. Details are specified on the article's page.

Mirzaei Danyal; HAJIZADEH MOHAMMAD; AZIZI ABDOLBASET; Koolivand Davoud

مرکز اطلاعات علمی Scientific Information Database (SID) - Trusted Source for Research and Academic Resources

Journal Paper

Paper Information

Journal: GENETIC ENGINEERING AND BIOSAFETY JOURNAL Year:2019 | Volume:8 | Issue:1 Page(s): 51-62

Download Full-Text

Persian Verion

View:

636

Download:

Cites:

Information Journal Paper

Title

Cloning and expression of Iranian isolate of Apple chlorotic leaf spot virus coat protein gene in Escherichia coli

Author(s)

Mirzaei Danyal | HAJIZADEH MOHAMMAD | AZIZI ABDOLBASET | Koolivand Davoud | Issue Writer Certificate

Keywords

Antibody

Cloning

Expression

Recombinant protein

SDS-PAGE

Abstract

The Apple chlorotic leafspot virus (ACLSV) is among of the most common virus infecting pome and stone fruit trees around the world. The virus has spread throughout the world and has been reported in many hosts such as; apples, pears, peaches and cherries. The purpose of this study is to express the coat protein (CP) of ACLSV in E. coli. CP Expression in E. coli has been developed to prepare antigen for Antibody production without needs to purify the virus from infected plant. In this study, complete CP gene (582 bp) from an isolate of ACLSV (SSa) was amplified by specific primers, MF and MR. The amplified CP gene was ligated to TA Cloning vector (pTG19-ACLSV CP) and transformed into E. coli strain DH5α . Transformed cells were selected on LB containing ampicillin, X-Gal and IPTG, and recombinant plasmids were confirmed by restriction analysis. Then, the pTGACLSV CP was sub-cloned into pET28a (+) as Expression vector that digested by BamHl and XhoI restriction enzymes. Finally, the pET28a-ACLSV CP was transformed into E. coli strain BL21 by heat shock method. For ACLSV CP Expression, the cells containing pET28a-ACLSV CP were induced by 1 mM of IPTG and the protein was extracted two and four hours after induction and analyzed in SDS-PAGE. The SDS-PAGE result showed that ACLSV CP have been expressed that is expected based on additional tags from plasmid.

Cites

No record.

References

No record.

Cite

APA: Copy

Mirzaei, Danyal, HAJIZADEH, MOHAMMAD, AZIZI, ABDOLBASET, & Koolivand, Davoud. (2019). Cloning and expression of Iranian isolate of Apple chlorotic leaf spot virus coat protein gene in Escherichia coli. GENETIC ENGINEERING AND BIOSAFETY JOURNAL, 8(1 ), 51-62. SID. https://sid.ir/paper/270550/en

Vancouver: Copy

Mirzaei Danyal, HAJIZADEH MOHAMMAD, AZIZI ABDOLBASET, Koolivand Davoud. Cloning and expression of Iranian isolate of Apple chlorotic leaf spot virus coat protein gene in Escherichia coli. GENETIC ENGINEERING AND BIOSAFETY JOURNAL[Internet]. 2019;8(1 ):51-62. Available from: https://sid.ir/paper/270550/en

IEEE: Copy

Danyal Mirzaei, MOHAMMAD HAJIZADEH, ABDOLBASET AZIZI, and Davoud Koolivand, “Cloning and expression of Iranian isolate of Apple chlorotic leaf spot virus coat protein gene in Escherichia coli,” GENETIC ENGINEERING AND BIOSAFETY JOURNAL, vol. 8, no. 1 , pp. 51–62, 2019, [Online]. Available: https://sid.ir/paper/270550/en

Related Journal Papers