The chicken leptin gene: Has it been cloned?
The chicken leptin gene: Has it been cloned?
复制标题
DOI：
10.1006/gcen.1999.7322
复制
发表时间：
1999-09-01
影响因子：
2.7
通讯作者：
Sharp, PJ
中科院分区：
文献类型：
--
作者：
Friedman-Einat, M;Boswell, T;Sharp, PJ
最近有报道称，鸡瘦素基因的DNA序列与小鼠瘦素序列具有95%的核苷酸相似性（Taouis et al., 1998, gene 208, 239-242）。为了证实这一发现，在两个独立的实验室里进行了实验。以小鼠瘦素序列为基础，设计了14条禽瘦素基因扩增引物。其中四个引物与小鼠和公布的鸡瘦素序列相同。对几种鸡品系的脂肪、肝脏和胰腺以及家火鹅和日本鹌鹑的基因组DNA和逆转录mRNA进行了PCR扩增。从禽类模板中没有得到与小鼠瘦素序列非常相似的PCR产物。当使用对照小鼠模板时，小鼠瘦素序列扩增一致。用小鼠瘦素探针对鸡脂肪和肝脏及鹅肥肝进行Northern杂交，均不能产生与poly(a)(+) RNA的杂交信号，而与对照小鼠脂肪总RNA的杂交信号较强。在低强度洗涤条件下进行南方杂交，发现小鼠瘦素探针与鸡基因组DNA杂交。在更严格的洗涤条件下，鸡的信号消失了，而来自对照小鼠和绵羊的基因组DNA则保留了下来。这表明假定的鸡瘦素序列在小鼠和绵羊基因之间的核苷酸序列同源性低于83%。由此可见，鸡基因组中不存在与小鼠瘦素序列相似的鸡瘦素基因序列。此外，家鸡、火鸡、鹅或日本鹌鹑的脂肪或肝脏中不存在与小鼠瘦素具有高度序列同一性的mRNA。(C) 1999学术出版社。
The DNA sequence of a chicken leptin gene that shares 95% nucleotide similarity with the mouse leptin sequence has been recently reported (Taouis et al., 1998, Gene 208, 239-242). Experiments have been performed independently in two laboratories to try to confirm this finding. Fourteen PCR primers based on the mouse leptin sequence were designed to amplify the avian leptin gene. Four of the primers were identical to the mouse and published chicken leptin sequences. PCR amplification was carried out on genomic DNA and reverse-transcribed mRNA from the fat, liver, and pancreas of several chicken strains and from the domestic turkey goose, and Japanese quail. No PCR products sharing close similarity to the mouse leptin sequence were generated from any avian templates. Amplification of mouse leptin sequence was consistently obtained when control mouse templates were used. Northern hybridization using a mouse leptin probe failed to produce a signal with poly(A)(+) RNA from chicken fat and liver and from the fat and liver of force-fed geese but a strong signal was obtained from control mouse fat total RNA. Southern hybridization under low stringency washing conditions revealed hybridization of a mouse leptin probe to chicken genomic DNA. Under higher stringency washing conditions, the chicken signal disappeared, while those from control mouse and sheep genomic DNA remained. This suggests that the putative chicken leptin sequence shares less than the 83% nucleotide sequence identity between the mouse and sheep genes. It is concluded that a chicken leptin gene sequence with close sequence similarity to mouse leptin is not present in the chicken genome. Furthermore, mRNA sharing high sequence identity with mouse leptin is not present in the fat or liver of the domestic chicken, turkey, goose, or Japanese quail. (C) 1999 Academic Press.