Construction and characterization of yeast two-hybrid cDNA library derived from LFBK cell line
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Title |
Construction and characterization of yeast two-hybrid cDNA library derived from LFBK cell line
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Creator |
Sonalika Mahajan
Gaurav Kumar Sharma Rakesh Matura Saravanan Subramaniam Jajati Keshari Mohapatra Bramhadev Pattnaik |
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Subject |
FMD, SMART, Yeast two-hybrid, cDNA library
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Description |
Not Available
The cDNA libraries are indispensable and critical tools for performing protein-protein interaction studies. In this study, a high quality yeast two-hybrid cDNA library from the LFBK cell line was constructed and characterized. LFBK cell line was originally derived from the swine kidney cells and is highly susceptible to foot-and-mouth disease virus (FMDV) infection. The total RNA was extracted from the LFBK cells and the switching mechanism at the 5' end of RNA template (SMART) technique was employed for the cDNA synthesis. Subsequently, double stranded cDNA was amplified by long-distance PCR, purified and co-transformed with pGADT7-rec vector in yeast strain Y187. The quality parameters of the constructed library were evaluated to qualify the constructed library. Nucleotide sequencing of the randomly selected clones from the library confirmed the swine genotype of LFBK cell line. The LFBK cDNA library was mated with the 2C protein of FMDV in yeast two-hybrid (YTH) system and several putative interaction partners were identified in the preliminary screening. The LFBK library was observed to be of high quality and could potentially be applied to protein interaction studies between FMDV and the host cells using YTH system. Not Available |
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Date |
2021-08-01T12:00:12Z
2021-08-01T12:00:12Z 2015-05 |
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Type |
Research Paper
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Identifier |
Mahajan S, Sharma GK, Matura R, Subramaniam S, Mohapatra JK, Pattnaik B. Construction and characterization of yeast two-hybrid cDNA library derived from LFBK cell line. Biologicals. 2015 May;43(3):202-8. doi: 10.1016/j.biologicals.2015.01.003. Epub 2015 Mar 16. PMID: 25787111.
Not Available http://krishi.icar.gov.in/jspui/handle/123456789/51828 |
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Language |
English
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Relation |
Not Available;
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Publisher |
Elsevier
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