Carboxyl-terminal sequence of entactin deduced from a cDNA clone.
Carboxyl-terminal sequence of entactin deduced from a cDNA clone.
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从 cDNA 克隆推断出巢蛋白的羧基末端序列。
DOI：
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发表时间：
1987
影响因子：
11.1
通讯作者：
A. Chung
中科院分区：
文献类型：
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作者：
M. Durkin;B. Carlin;J. Vergnes;B. Bartos;J. Merlie;A. Chung
Enlavin是一种广泛分布的基底膜硫酸化糖蛋白，分子量约为150 kDa。巢蛋白基因在小鼠胚胎发育早期表达。通过抗体筛选构建在λ gt 11表达载体中的oligo（dT）-primed cDNA文库，分离出两个与大鼠巢蛋白mRNA互补的cDNA克隆。将其中一个克隆λ 1 E亚克隆到质粒pBR 322中并进一步表征。该克隆含有与长度为6个内切酶的mRNA种类互补的序列。该mRNA在兔网织红细胞裂解物中翻译，产生143 kDa的多肽，该多肽用抗巢蛋白抗血清沉淀。对长1328个碱基对的cDNA插入片段进行测序，发现其包含729个碱基对的开放阅读框，编码巢蛋白羧基末端的243个氨基酸。肽的分析显示没有延伸的α-螺旋或β-折叠二级结构。用pLambda 1 E缺口翻译法制备的放射性标记探针监测F9胚胎癌细胞中巢蛋白mRNA的稳态水平，F9胚胎癌细胞通过暴露于视黄酸和二丁酰环AMP诱导分化。巢蛋白mRNA的稳态水平的增加滞后于层粘连蛋白B2链mRNA的增加，表明层粘连蛋白和巢蛋白是独立的，而不是协调调节。
Entactin is a widely distributed basement membrane sulfated glycoprotein of approximately equal to 150 kDa. The entactin gene is expressed early in mouse embryogenesis. Two cDNA clones complementary to rat entactin mRNA were isolated by antibody screening of an oligo(dT)-primed cDNA library constructed in the lambda gt11 expression vector. One of the clones, lambda 1E, was subcloned into plasmid pBR322 and further characterized. The clone contained sequences complementary to an mRNA species 6 kilobases in length. This mRNA was translated in rabbit reticulocyte lysates to yield a polypeptide of 143 kDa that was precipitated with anti-entactin antiserum. The cDNA insert, 1328 base pairs long, was sequenced and found to contain an open reading frame of 729 base pairs that coded for 243 amino acids at the carboxyl terminus of entactin. Analysis of the peptide revealed no extended alpha-helical or beta-sheet secondary structures. Radiolabeled probes prepared by nicktranslation of p lambda 1E were used to monitor the steady-state levels of entactin mRNA in F9 embryonal carcinoma cells that were induced to differentiate by exposure to retinoic acid and dibutyryl cyclic AMP. The increase in steady-state levels of entactin mRNA lagged behind the increase in mRNA for the B2 chain of laminin, suggesting that laminin and entactin are independently rather than coordinately regulated.