Expression Of The Human Soluble Low Density Lipoprotein Receptor (sLDLR) In Pichina Pastoris And Study Of The Screening Model For Low Density Lipoprotein Receptor Antagonist

Low density lipoprotein receptor (LDLR) plays important role in regulation of cholesterol homeostasis. Under the influence by various factors, the function of LDLR is reduced or deleted. As result, the person will suffer from atherosclerosis. LDLR may be as target for screening new drugs for anti-atherosclerosis. In this paper, we reported that the sLDLR gene for extracellular Ligand binding domain was expressed in Pichia pastoris GS115. A screening model for LDLR antagonists has been constructed. Using RT-PCR, the sLDLR cDNA was amplified with the total RNA of human hepatoma Bel- 7402 as template. The nucleotide sequencing analysis for sLDLR cDNA is same with reported except a synonym mutation. The sLDLR cDNA was inserted the downstream of α-signal peptide encoding gene in the plasimd pPIC9K. After linearization by restriction enzyme digestion, the recombinant plasmid was transformated into Pichia pastoris GS115 with electroporation.For His~+ transformants, the Mut~S phenotype colony growed slowly on the MM medium were selected. After identification with PCR, the Mut~S...