Construction and identification of yeast cDNA expression library of human dermal papillae cells in primary culture

VernacularTitle:原代培养人毛乳头细胞cDNA酵母表达文库的构建和鉴定
Author: Zhiqiang SONG ; Lihua SUN ; Zhifang ZHAI ; Fei HAO
Publication Type:Journal Article
Keywords: Hair; Dermal papilla; Two-hybrid system techniques; Gene library
From: Chinese Journal of Dermatology 2010;43(6):408-410
CountryChina
Language:Chinese
Abstract: Objective To construct yeast cDNA expression library of human dermal papillae cells (DPCs) in primary culture.Methods Human dermal papilla cells (DPCs) were isolated by two-step digestion method and cultured in DMEM medium.Total RNA was extracted from primary DPCs that exhibited an aggregative behavior in culture,then,cDNA was synthesized and amplified by using CloneMinerTM cDNA Library Construction kit to construct primary cDNA library and yeast cDNA expression libary.Results The average titer and total clones were 7.0×106 colony forming units(cfu)/ml and 1.4×107 cfu respectively in the primary library,5.5×106 cfu/ml and 1.1×107 cfu respectively in the yeast expression library.The average insert size Was 1.2 kb and the recombination rate was above 95%.Conclusions The yeast cDNA expression library of DPCs in primary culture has been successfully constructed.which will lay a foundation for screening proteins interacting with HSPC016 gene in DPCs with yeast two-hybrid system.