Anathy V, Venugopal T, Koteeswaran R, Pandian T J, Mathavan S
Department of Genetics, School of Biological Sciences, Madurai Kamaraj University, Madurai 625 021, India.
J Biosci. 2001 Sep;26(3):315-24. doi: 10.1007/BF02703740.
A tissue-specific cDNA library was constructed using polyA+ RNA from pituitary glands of the Indian catfish Heteropneustes fossilis (Bloch) and a cDNA clone encoding growth hormone (GH) was isolated. Using polymerase chain reaction (PCR) primers representing the conserved regions of fish GH sequences the 3' region of catfish GH cDNA (540 bp) was cloned by random amplification of cDNA ends and the clone was used as a probe to isolate recombinant phages carrying the full-length cDNA sequence. The full-length cDNA clone is 1132 bp in length, coding for an open reading frame (ORF) of 603 bp; the reading frame encodes a putative polypeptide of 200 amino acids including the signal sequence of 22 amino acids. The 5' and 3' untranslated regions of the cDNA are 58 bp and 456 bp long, respectively. The predicted amino acid sequence of H. fossils GH shared 98% homology with other catfishes. Mature GH protein was efficiently expressed in bacterial and zebrafish systems using appropriate expression vectors. The successful expression of the cloned GH cDNA of catfish confirms the functional viability of the clone.
利用印度鲶鱼（Heteropneustes fossilis，布洛赫）垂体的聚腺苷酸加尾RNA构建了组织特异性cDNA文库，并分离出一个编码生长激素（GH）的cDNA克隆。使用代表鱼类GH序列保守区域的聚合酶链反应（PCR）引物，通过cDNA末端的随机扩增克隆了鲶鱼GH cDNA的3'区域（540 bp），并将该克隆用作探针来分离携带全长cDNA序列的重组噬菌体。全长cDNA克隆长度为1132 bp，编码一个603 bp的开放阅读框（ORF）；该阅读框编码一个推定的200个氨基酸的多肽，包括22个氨基酸的信号序列。cDNA的5'和3'非翻译区分别长58 bp和456 bp。印度鲶鱼GH的预测氨基酸序列与其他鲶鱼的同源性为98%。使用合适的表达载体，成熟的GH蛋白在细菌和斑马鱼系统中得到了有效表达。鲶鱼克隆的GH cDNA的成功表达证实了该克隆的功能活性。
