为克隆并分析小鼠催乳素释放肽基因, 从小鼠下丘脑提取总RNA, 进行RT-PCR, PCR产物与pMD-19T连接后转化E.coli DH5α, 检测阳性克隆并测序。测序结果表明, 所得序列与大鼠催乳素释放肽编码区序列的相似指数为86.3%。克隆所得序列为小鼠催乳素释放肽的基因片段。
To clone and analyse mouse prolactin-releasing peptide. Total RNA was extracted from hypothalamus and amplified by RT-PCR. The PCR products were ligated into the pMD-19T vector, and then transformed into competent cells of E. coli DH5a, The positive cloning was sequenced and anelyzed.The cloned nucleotide sequence shared 86.3% hourology with the code zone sequence of rat prolactin-releasing peptide. The cloned nucleotide sequence was pert of mouse polactin-releasing peptide gene.
[1] Shuji Hinuma, Yugo Habata, Ryo Fujiii, et al. A Prolactin-releasing peptide in zhe brain[J]. Nature, 1998, 393: 273-276.
[2] Maruyama M, Matsumoto H, Fujiwara K, et al. Immuno cytochemical localization of prolactin-releasing peptide in the rat brain[J]. Endocrinology, 1999, 140(5): 2326-2333.
[3] Fujiwara K, Matsumoto H, Yada T, et al. Identification of the prolactin-releasing peptide-producing cell in the rat adrenal gland[J]. Regul Pept 2005, 126(1-2): 97-102.
[4] Fujii R, Fukusumi S, Hosoya M, et al. Tissue distribution of prolactin-releasing peptide(PrRP) and its receptor[J]. Regul Pept 1999, 83(1): 1-10.
[5] Vergoni A V, Watanobe H, Guidetti G, et al. Effect of repeated administration of prolactin releasing peptide on feeding behavior in rats[J]. Brain Res, 2002, 955(1-2): 207-213.
[6] Zhang SQ, Inoue S, Kimura M, et al. Prolactin-releasing peptide is a potent mediator of stress responses in the brain through the hypothalamic paraventricular nucleus[J]. Neuroscience, 2006, 141(2): 1069-1086.
[7] Zhang SQ, Inoue S, Kimura M. Sleep-promoting activity of prolactin-releasing peptide(PrRP) in the rat[J]. Neuroreport, 2001, 12(15): 3173-3176.
[8] Masanobu Yamada, Atsushi Ozawa, Sumiyasu Ishii, et al. Isolation and Characterization of the Rat Prolactin-Releasing Peptide Gene: Multiple TATA Boxes in the Promoter Region[J]. Biochemical and Biophysical Research Communications, 2001, 281(1): 53-56
[9] Curlewis J D, Kusters D H, Barclay J L, et al. Prolactin-releasing peptide in the ewe: cDNA cloning, mRNA distribution and effects on prolactin secretion in vitro and in vivo[J]. J Endocrinol, 2002, 174(1): 45-53.
[10] Fukusumi S, Fujii R, Hinuma S. Recent advances in mammalian RFamide peptides: the discovery and functional analyses of PrRP, RFRPs and QRFP[J]. Peptides, 2006, 27(5): 1073-1086.
