Partial titin cDNA sequence isolated from rabbit cardiac muscle RNA.
Partial titin cDNA sequence isolated from rabbit cardiac muscle RNA.
复制标题
从兔心肌 RNA 中分离出部分肌联蛋白 cDNA 序列。
DOI：
10.1007/bf00123100
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发表时间：
1993
影响因子：
2.7
通讯作者：
Greaser,ML
中科院分区：
文献类型：
--
作者：
Fritz,JD;Wolff,JA;Greaser,ML
使用基于兔骨骼肌肌联蛋白（连接蛋白）cDNA 的引物，从兔心肌总 RNA 中扩增兔心脏肌联蛋白 cDNA 的两个区域。这些 1.7 kb 和 1.5 kb RNA-PCR 产物分别基于骨骼肌肌联蛋白克隆 CE12 和 MS2 的 3' 区域。通过在 RNA-PCR 中使用随机引物的新型 3' 延伸技术，将 1.7 kb 产物的 cDNA 序列额外延伸 1.5 kb。心脏肌联蛋白 cDNA 与骨骼肌对应物的核苷酸序列有 99% 相同，并预测有两种类型的 100 个残基重复序列，Southern 印迹分析表明心脏和骨骼肌肌联蛋白均由同一基因编码。使用肌联蛋白特异性引物对人基因组 DNA 进行 PCR 扩增，表明兔和人肌联蛋白序列之间存在很强的序列相似性。从人类基因组 DNA 中成功扩增 907 个碱基对区域表明，肌联蛋白含有跨越多个基序边界的大外显子。这对于处理如此大的 RNA 转录本可能特别有利。
Two regions of the rabbit cardiac titin cDNA were amplified from rabbit cardiac muscle total RNA using primers based on rabbit skeletal muscle titin (connectin) cDNAs. These 1.7 kb and 1.5 kb RNA-PCR products were based on the 3′ regions of the skeletal muscle titin clones CE12 and MS2, respectively. The cDNA sequence of the 1.7 kb product was extended an additional 1.5 kb by a novel 3′ extension technique which used random primers in RNA-PCR. The cardiac titin cDNAs were 99% identical in nucleotide sequence to their skeletal muscle counterparts and predicted two types of 100-residue repeats, Southern blot analysis suggested that both cardiac and skeletal titin are encoded by the same gene. PCR amplification of human genomic DNA with titin specific primers indicated that there is strong sequence similarity between rabbit and human titin sequences. The successful amplification of a 907 basepair region from human genomic DNA suggested that titin contains large exons which span multiple motif borders. This may be particularly advantageous in the processing of such a large RNA transcript.