Ho W K, Tsang W H, Dias N P
Biotechnology Laboratory, Chinese University of Hong Kong.
Biochem Biophys Res Commun. 1989 Jun 30;161(3):1239-43. doi: 10.1016/0006-291x(89)91375-2.
We have constructed a cDNA library in lamda gt11 using mRNA isolated from the pituitary glands of the grass carp (Ctenopharyngodon idellus). Based on the published sequence of the rainbow trout growth hormone cDNA, we synthesized two oligonucleotide probes. One of these hybridized strongly with a specific mRNA fragment from the grass carp pituitary. Using this probe, we have isolated six positive clones carrying an insert of approximately 1.2 Kb. By restriction enzyme digestion, all the clones were determined to be identical. Sequence determination on one of them indicated that it has an open reading frame coding for 210 amino acids. Both the nucleotide and translated amino acid sequence are highly homologous to those of the salmon growth hormone and the common carp. A putative signal peptide consisting of hydrophobic amino acids can be identified at the 5' end of the sequence. A polyadenylation signal, ATTAAA, was also present 12 base upstream from the poly A tail.
我们使用从草鱼(Ctenopharyngodon idellus)垂体中分离的mRNA,构建了一个λgt11 cDNA文库。基于已发表的虹鳟生长激素cDNA序列,我们合成了两个寡核苷酸探针。其中一个与草鱼垂体中的一个特定mRNA片段强烈杂交。使用这个探针,我们分离出了六个携带约1.2 Kb插入片段的阳性克隆。通过限制性酶切分析,确定所有克隆都是相同的。对其中一个克隆进行序列测定表明,它有一个编码210个氨基酸的开放阅读框。核苷酸序列和翻译后的氨基酸序列与鲑鱼生长激素和鲤鱼的序列高度同源。在序列的5'端可以鉴定出一个由疏水氨基酸组成的假定信号肽。在poly A尾上游12个碱基处也存在一个聚腺苷酸化信号ATTAAA。