利用锌指基序的保守性从人脑组织分离新的C_2H_2型锌指蛋白基因表达序列  

ISOLATION OF NOVEL EXPRESSION SEQUENCES OF C_2H_2 TYPE ZINC FINGER PROTEIN GENE FROM HUMAN BRAIN TISSUE ACCORDING TO THE CONSERVATION OF ZINC FINGER MOTIF

作　　者：孙喜元[1] 余龙[1] 吴国俊[1] 范玉新[1] 郑其平[1] 胡培蓉[1] 张民[1] 江萤 刘澍[1] 许月芳 赵寿元[1] 

机构地区：[1]遗传工程国家重点实验室,复旦大学遗传学研究所,上海200433

出　　处：《实验生物学报》1998年第4期377-382,共6页Acta Biologiae Experimentalis Sinica

基　　金：国家自然科学基金(批准号:39525015;39680019);国家863高技术项目资助~~

摘　　要：本文利用锌指基序的保守性设计引物,在低严谨条件下扩增人基因组总DNA,获得8个长度呈梯度的PCR扩增产物电泳条带。取其中第2和第3电泳条带,回收DNA片段并将它们克隆,经测序和查新比较,共获得60个含有锌指蛋白基因基序的单一的序列,其中23个为新的锌指蛋白基因DNA片段。以它们为探针,杂交筛选人脑组织cDNA文库,得到初筛cD-NA克隆44个。对其中28个初筛cDNA克隆进行复筛之后,得到20个cDNA单克隆。对这些阳性克隆进行测序,读出18个含有锌指蛋白基因基序的序列,国际联网查新之后,证明其中16个是新的锌指蛋白基因片段。这些新的锌指蛋白基因片段为今后克隆有意义的全长锌指蛋白cDNA提供了重要的实验材料。By low stringency PCR amplification of ge-nomic DNA using the primers designed based on the conservation of zinc finger motif, we got 8 gradient eletrophoretic bands. After recovery of the second and third bands, the DNA fragments in them were cloned and sequenced. Compared to the GenBank database, among these 60 segments containing zinc finger motif, 23 segments were novel zinc finger genes' genomic segments. Then the human brain tissue cDNA library was screened, using these segments as probes, and44 positive clones were obtained. Rescreening 28 of them, we got 20 rescreened clones. All of them were sequenced and sent to the GenBank DNA database for sequence analysis, the results showed that 16 were novel C2H2 type zinc finger protein cDNA segments. The cDNA segments encoding the novel C2H2 type zinc finger proteins provide the basic materials for cloning of full length cDNA of valuable novel zinc finger protein genes.

关 键 词：人 C2H2型锌指蛋白 基因 表达序列 

分 类 号：Q523[生物学—生物化学] Q987