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- Title
Buffalo Leukemia Inhibitory Factor Induces Differentiation and Dome-Like Secondary Structures in COS-1 Cells.
- Authors
Kaur, Gurjeet; Ali, Syed Azmal; Pachauri, Shikha; Malakar, Dhruba; Kaushik, Jai K.; Mohanty, Ashok K.; Kumar, Sudarshan
- Abstract
This study aimed to understand the molecular characteristics of buffalo leukemia inhibitory factor (BuLIF) and the generation of a stably transfected COS-1_BuLIF cell line for its functional characterization. Cumulus cells, isolated from oocytes, were separated, and total cDNA was prepared. The BuLIF gene was ligated into the cloning vector pJET1.2/blunt and expression vector pAcGFP-N1 which was transfected into COS-1 cells and confirmed by qRT-PCR and Western blot. BuLIF was immunoprecipitated and evaluated through a MTT assay. qRT-PCR of STAT3 was performed. The multiple sequence alignment of BuLIF showed high similarity with sheep (98.77%) and cattle (96.62%) compared with other species. The BuLIF gene has an open reading frame of 609 nucleotides coding for 202 amino acids. BuLIF was integrated into the genome of COS-1 cells and resulted in the formation of dome-like secondary structures which are indicative of its functional role mediated through STAT3 proteins. In conclusion, this cell line is suitable for understanding LIFmediated biological functions.
- Subjects
ANIMAL models of leukemia; ANTISENSE DNA; IMMUNOPRECIPITATION; POLYMERASE chain reaction; CELL lines
- Publication
Cytogenetic & Genome Research, 2017, Vol 151, Issue 3, p119
- ISSN
1424-8581
- Publication type
Article
- DOI
10.1159/000465507