人基质细胞中新基因的获得及目的基因的克隆  

Obtaining of Three New ESTs from Human Fetal Bone Marrow Stromal Cells cDNA Expression Library and Screening of Target Genes

作　　者：黄玉珊[1] 赵贵军[1] 吴文言[1] 徐安龙[1] 

机构地区：[1]中山大学生命科学学院,广东广州510275

出　　处：《中山大学学报（自然科学版）》2001年第5期88-90,共3页Acta Scientiarum Naturalium Universitatis Sunyatseni

基　　金：广东省自然科学基金资助项目(970612)

摘　　要：取连续培养的骨髓基质细胞并提取其ｍＲＮＡ，合成ｃＤＮＡ后，收集＞４００ｂｐ的片段，连接到真核表达载体ｐｃＤＮＡ３．０上，构建人胎儿骨髓基质细胞真核ｃＤＮＡ质粒文库，并以ＰＣＲ的方法从文库中扩增出编码人ＩＬ－６、ＳＣＦ的基因序列，随机对文库克隆进行测序，得到了３个新基因ＥＳＴ，其中２个在ＧｅｎＢａｎｋ的登录号分别为ＡＦ２４４９９８及ＡＦ２４４９９９．mRNA were isolated from continuously cultured human fetal bone marrow stromal cells and transcripted into cDNA. Fragments (>400 bp) were obtained through sizesep 400 columns, then ligated into eukaryon expres-sion plasmid pcDNA3 .0. Three new ESTs were found through sequencing 15 library clones, they are quite different-ly from the sequences in GenBank. Maybe those genes are stromal specific genes. Full length cDNA encoding hu-man stem cell factor and IL -6 were amplified by PCR using this library as templates.

关 键 词：骨髓基质细胞 CDNA文库 EST 测序 基因 克隆 

分 类 号：Q78[生物学—分子生物学]