人前列腺癌细胞系PC-3中PTI-1 cDNA的克隆及序列分析

为获取人前列腺癌诱导基因家族新成员 ,以人前列腺癌细胞系PC 3的总RNA为模板 ,用RT PCR方法扩增人前列腺癌诱导基因 1(PTI 1) ,将其亚克隆至克隆载体pUC19,进行测序 .将克隆得到的PTI 1基因片段与国外报道的人PTI 1基因编码区cDNA同源性为 99.3% ,但第 72 6、74 6、116 7、12 70、132 6、144 9、16 94和 16 95位碱基分别由A、A、C、C、A、A、T和C替代了文献中的C、C、G、G、G、C、C和T .其中 6个碱基的不同 ,导致了第 36、183、2 17、2 36、2 77和 35 9位密码子代表的氨基酸分别由Lys替代Gln ,Arg替代Gly ,Ala替代Gly ,Ser替代Gly ,Thr替代Pro和Arg替代Cys .将所获得的基因在GenBank登录 ,登录号为AF3974 0 3.结果提示 ,获得了PTI 1家族新成员 ,并且与EF 1α分子的同源性较已报道的PTI 1高 .

Cloning and Sequencing of PTI-1 cDNA in Human Prostate Carcinoma Cell Line PC-3

To obtain a new member of the human prostate carcinoma tumor inducing gene ( PTI 1) family in human prostate carcinoma cell line PC 3, human PTI 1 cDNA was synthesized from human prostate carcinoma cell line PC 3 by RT PCR, which was then subcloned into pUC19 vector and sequenced. The data indicate that there were eight bases substitutions(726, 7446, 1167, 1326, 1449, 1695) leading subsequently to the change of six amino acids(36, 183, 217, 236, 277, 359) compared to that previously reported. Thus, Lys, Arg, Ser, Thr and Arg were substituted by Gln, Gly, Gly, Gly, Pro and Cys, respectively. The cDNA sequence of PTI 1 was registered in GenBank with accession number AF 397403, A new member of PTI 1 family was sequenced and its homology with EF 1 α was even higher than that previously reported.