ALTERNATIVE SPLICING OF HUMAN ELASTIN MESSENGER-RNA INDICATED BY SEQUENCE-ANALYSIS OF CLONED GENOMIC AND COMPLEMENTARY-DNA
ALTERNATIVE SPLICING OF HUMAN ELASTIN MESSENGER-RNA INDICATED BY SEQUENCE-ANALYSIS OF CLONED GENOMIC AND COMPLEMENTARY-DNA
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DOI：
10.1073/pnas.84.16.5680
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发表时间：
1987-08-01
影响因子：
11.1
通讯作者：
ROSENBLOOM, J
中科院分区：
文献类型：
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作者：
INDIK, Z;YEH, H;ROSENBLOOM, J
从单个7月龄胎儿人主动脉中分离的Poly（A）+ RNA用于通过RNase H方法合成cDNA，并将cDNA插入到Gt10中。含有弹性蛋白序列的重组噬菌体通过与克隆的、含有人弹性蛋白基因外显子的片段杂交来鉴定。选择含有3.3、2.7和2.3内切酶插入片段的三个克隆用于进一步分析。还从基因组文库中分离出三个含有17.8个人弹性蛋白酶基因的重叠克隆。对六个克隆的完整序列分析表明：（i）cDNA包含编码786个氨基酸的mRNA的整个翻译部分，包括先前在猪原弹性蛋白中未鉴定的几个不常见的亲水性氨基酸序列，（ii）编码蛋白质中的疏水或交联结构域的外显子在基因中交替，和（iii）大量的Alu重复序列出现在整个内含子中。数据还表明mRNA的大量选择性剪接。这些结果表明，在人群中的弹性纤维的精确分子结构的显着变化的潜力。
Poly(A)+ RNA, isolated from a single 7-mo fetal human aorta, was used to synthesize cDNA by the RNase H method, and the cDNA was inserted into .lambda.gt10. Recombinant phage containing elastin sequences were identified by hybridization with cloned, exon-containing fragments of the human elastin gene. Three clones containing inserts of 3.3, 2.7, and 2.3 kilobases were selected for further analysis. Three overlapping clones containing 17.8 kilobases of the human elastin gene were also isolated from genomic libraries. Complete sequence analysis of the six clones demonstrated that: (i) the cDNA encompassed the entire translated portion of the mRNA encoding 786 amino acids, including several unusual hydrophilic amino acid sequences not previously identified in porcine tropoelastin, (ii) exons encoding either hydrophobic or crosslinking domains in the protein alternated in the gene, and (iii) a great abundance of Alu repetitive sequences occurred throughout the introns. The data also indicated substantial alternative splicing of the mRNA. These results suggest the potential for significant variation in the precise molecular structure of the elastic fiber in the human population.