Freyer G A, Robbins J
J Biol Chem. 1983 Jun 10;258(11):7149-54.
A cDNA library has been constructed in the plasmid pBR322 using a large size class of RNA derived from chicken embryonic leg muscle as the template material. A clone containing a 2350-base pair insert was selected and identified as coding for the myosin heavy chain sequence, based upon its ability to hybridize to genomic myosin heavy chain clones, and by direct nucleotide sequencing. Cross-hybridization experiments with myosin heavy chain genomic clones, and mRNAs derived from different muscle types were used to explore the heterogeneity of the various myosin heavy chain isoforms at the level of the coding sequences. Although extensive sequence homology with the other isoforms was observed, a fast white isoform-specific subclone was constructed, and used to demonstrate that different genes code for the adult and embryonic fast white myosin heavy chain proteins.
利用从鸡胚腿部肌肉中提取的一大类RNA作为模板材料，在质粒pBR322中构建了一个cDNA文库。选择了一个含有2350个碱基对插入片段的克隆，并根据其与基因组肌球蛋白重链克隆杂交的能力以及直接核苷酸测序，确定其编码肌球蛋白重链序列。利用肌球蛋白重链基因组克隆与来自不同肌肉类型的mRNA进行交叉杂交实验，在编码序列水平上探索各种肌球蛋白重链同工型的异质性。尽管观察到与其他同工型有广泛的序列同源性，但构建了一个快速白色同工型特异性亚克隆，并用于证明不同的基因编码成年和胚胎快速白色肌球蛋白重链蛋白。
