Suppr超能文献

狒狒脂蛋白脂肪酶:两种转录本的cDNA序列及可变的组织特异性表达

Baboon lipoprotein lipase: cDNA sequence and variable tissue-specific expression of two transcripts.

作者信息

Cole S A, Hixson J E

机构信息

Southwest Foundation for Biomedical Research, San Antonio, TX 78228-0147, USA.

出版信息

Gene. 1995 Aug 19;161(2):265-9. doi: 10.1016/0378-1119(95)00301-l.

Abstract

We have isolated two lipoprotein lipase (LPL)-encoding cDNA (LPL) clones from a baboon cardiac cDNA library, one of which spans a region from nucleotide (nt) 705 of the coding sequence to the poly(A) tail (2.8 kb). We used reverse transcription followed by PCR (RT-PCR), and anchor-ligated rapid amplification of cDNA ends (RACE) to amplify the remaining 5' region of the LPL transcript. Sequence comparisons reveal that the baboon nt sequence is 95% identical to the human cDNA sequence (ranging from 97.5 to 92.7% in the coding and noncoding regions, respectively). Less than 2% of nt substitutions cause changes between baboon and human amino acid (aa) sequences. The aa in the catalytic triad residues, the heparin-binding site in exon 6, as well as aa in positions where missense mutations cause LPL deficiency, are identical in baboons and humans. Characterization of the tissue-specific expression of LPL using Northern blots of total RNA showed that spinal cord expressed the most LPL transcripts of all baboon tissues examined. Like humans, baboons have two transcript sizes of approx. 3.6 and 3.4 kb in most tissues that express LPL, and sequencing of the 3' untranslated region (UTR) shows this is due to two polyadenylation sites. In contrast, only the larger 3.6-kb transcript is detected in RNA isolated from central nervous system (CNS) tissues. We used RT-PCR to show that the polyadenylation signal that produces the 3.4-kb message is present in CNS LPL transcripts, but is not utilized.

摘要

我们从狒狒心脏cDNA文库中分离出两个脂蛋白脂肪酶(LPL)编码cDNA(LPL)克隆,其中一个跨越编码序列的核苷酸(nt)705至聚腺苷酸尾(2.8 kb)区域。我们采用逆转录后进行PCR(RT-PCR)以及锚定连接的cDNA末端快速扩增(RACE)来扩增LPL转录本其余的5'区域。序列比较显示,狒狒的nt序列与人类cDNA序列的同一性为95%(编码区和非编码区分别为97.5%至92.7%)。狒狒和人类氨基酸(aa)序列之间因nt替换导致的变化不到2%。催化三联体残基中的aa、外显子6中的肝素结合位点以及错义突变导致LPL缺乏的位置上的aa,在狒狒和人类中是相同的。使用总RNA的Northern印迹法对LPL的组织特异性表达进行表征,结果表明在所有检测的狒狒组织中,脊髓表达的LPL转录本最多。与人类一样,在大多数表达LPL的组织中,狒狒有两种转录本大小,约为3.6 kb和3.4 kb,对3'非翻译区(UTR)进行测序表明,这是由于两个聚腺苷酸化位点所致。相比之下,在从中枢神经系统(CNS)组织分离的RNA中仅检测到较大的3.6 kb转录本。我们使用RT-PCR表明,产生3.4 kb信息的聚腺苷酸化信号存在于CNS的LPL转录本中,但未被利用。

文献检索

告别复杂PubMed语法,用中文像聊天一样搜索,搜遍4000万医学文献。AI智能推荐,让科研检索更轻松。

立即免费搜索

文件翻译

保留排版,准确专业,支持PDF/Word/PPT等文件格式,支持 12+语言互译。

免费翻译文档

深度研究

AI帮你快速写综述,25分钟生成高质量综述,智能提取关键信息,辅助科研写作。

立即免费体验