[Cloning and expression of pituitary prolactin gene in Ailuropoda melanoleuca].

Autor: Zheng X; Institute of Genetics, College of Life Sciences, Zhejiang University, Hangzhou 310012, China. luciferase@21cn.com, Zhang ZH, Hu XL, Liao MJ, Zhang AJ, Zhu MY
Jazyk: čínština
Zdroj: Yi chuan xue bao = Acta genetica Sinica [Yi Chuan Xue Bao] 2004 Sep; Vol. 31 (9), pp. 963-9.
Abstrakt: The giant panda (Ailuropoda melanoleuca) is an endangered species and indigenous to China. It has been proposed that it has a highly specialized reproductive pattern with low fecundity, but little is known about its basic reproductive biology at molecular level. In this study,the pituitary prolactin (PRL) cDNA of giant panda was amplified by RT-PCR from pituitary total RNA and then cloned, sequenced and submitted to GenBank (GenBank accession No. AY161285). The sequence analysis revealed that the giant panda prolactin cDNA contains a 687-nucleotide open reading frame encoding the prolactin prohormone of 229 amino acid residues. The signal peptide contains 30 amino acid residues and the mature prolactin is composed of 199 amino acid residues. Then the DNA fragment amplified was subcloned into pGEX-4T-1 procaryotic expression plasmid and protein expression was induced by IPTG in Escherichia coil BL21. SDS-PAGE analysis revealed the PRL protein is infusible. The multiple sequence alignments revealed that the homology of giant panda is 95% to cat and pig, 80% - 70% to human, cow and goat, 52% to rat and 45.9% to mouse at the amino acid level. The 64th amino acid of giant panda prolactin is hydrophilic serine instead of hydrophobic proline of cat, goat, and cow or hydrophobic alanine of human.
Databáze: MEDLINE