Fung M R, Campbell R M, MacGillivray R T
Nucleic Acids Res. 1984 Jun 11;12(11):4481-92. doi: 10.1093/nar/12.11.4481.
Thirty thousand colonies of a bovine liver cDNA library were screened with a mixture of synthetic oligodeoxyribonucleotides coding for bovine factor X. Five positive colonies were identified, and plasmid DNA was isolated. Cleavage with restriction endonucleases showed that these plasmids (designated pBX1 -5) contained inserts of 1530bp , 770bp , 700bp , 1100bp and 930bp . DNA sequence analysis of the plasmid with the largest insert ( pBX1 ) confirmed that bovine factor X cDNAs had been cloned. The cDNA sequence predicts that factor X is synthesized as a single chain precursor in which the light and heavy chains of plasma factor X are linked by the dipeptide Arg-Arg. The cDNA sequence also predicts that factor X is synthesized with a prepro leader peptide. We propose that at least five specific proteolytic events occur during the conversion of prepro -factor X to plasma factor Xa.
用编码牛凝血因子X的合成寡脱氧核糖核苷酸混合物筛选了三万个牛肝脏cDNA文库菌落。鉴定出五个阳性菌落,并分离出质粒DNA。用限制性内切酶切割表明,这些质粒(命名为pBX1 - 5)含有1530bp、770bp、700bp、1100bp和930bp的插入片段。对插入片段最大的质粒(pBX1)进行DNA序列分析,证实已克隆到牛凝血因子X的cDNA。cDNA序列预测,凝血因子X作为单链前体合成,其中血浆凝血因子X的轻链和重链由二肽精氨酸-精氨酸连接。cDNA序列还预测,凝血因子X是与前原导肽一起合成的。我们认为,在前原凝血因子X转化为血浆凝血因子Xa的过程中至少发生了五个特定的蛋白水解事件。