Molecular cloning and characterization of partial promoter of Acetyl coenzyme A carboxylase B gene in chicken

Poultry industry has achieved its present position from a mere backyard venture mainly due to selection for rapid early growth, optimum nutrition and rearing environment. However, intensive genetic selection for growth inadvertently selected for increased carcass fat. Hence, study was undertaken to identify partial promoter sequence and expression profile of ACACB gene as it mainly responsible for fat synthesis. An upstream part of ACACB gene with 697bp fragment was chosen and cloned in to a promoter less pAcGFP1-1 vector. The recombinant-vector was transfected in to cell culture of chicken embryo fibroblast cells. After two days of transfection, total RNA, DNA and protein were isolated and examined for expression of reporter gene. The 697bp fragment displayed a considerable amount of fluorescence under fluorescent microscope along with preferred band of 250bp of GFP on gel. The mRNA expression depicted the magnitudes of ∆C_t of 19.92 in transfected cells where there was no expression of GFP in control cells for GFP gene. The expression was also checked in western blot having presence of GFP in transfected cells. It is concluded that 697bp upstream fragment of ACACB gene had potential to be the promoter to regulate gene expression in the in-vivo cell culture medium.