Hossle J P, Rosenberg U B, Schäfer B, Eppenberger H M, Perriard J C
Nucleic Acids Res. 1986 Feb 11;14(3):1449-63. doi: 10.1093/nar/14.3.1449.
cDNA clones for chicken B-CK were isolated by immunoscreening from a gizzard cDNA library constructed in the expression vector lambda gtll. The entire coding portion in addition to the complete 3' untranslated region and 42 bp of the 5' noncoding part are represented in the clone H4. On RNA blots H4 insert DNA hybridized to a 1600 bp poly(A)+ RNA from gizzard, brain and heart but not to breast or skeletal muscle RNA. In vitro generated sense strand transcripts of H4 insert DNA were translated in vitro into a protein indistinguishable from isolated, authentic B-CK. The distinct nucleotide sequences of H4 insert DNA and M-CK cDNA were translated into 82% homologous amino acid sequences. Sequence heterogeneity among the B-CK cDNA clones within both the 3' noncoding and even in the coding region indicates the existence of multiple B-CK mRNA species.
通过免疫筛选,从构建于表达载体λgtll中的鸡肌胃cDNA文库中分离出鸡B-CK的cDNA克隆。克隆H4包含整个编码区,以及完整的3'非翻译区和5'非编码区的42个碱基对。在RNA印迹上,H4插入DNA与来自肌胃、脑和心脏的1600 bp聚腺苷酸加尾RNA杂交,但不与乳腺或骨骼肌RNA杂交。体外转录产生的H4插入DNA有义链转录本在体外被翻译成一种与分离得到的天然B-CK无法区分的蛋白质。H4插入DNA和M-CK cDNA的不同核苷酸序列被翻译成82%同源的氨基酸序列。3'非编码区甚至编码区内的B-CK cDNA克隆之间的序列异质性表明存在多种B-CK mRNA种类。