Degen S J, Davie E W
Children's Hospital Research Foundation, Cincinnati, Ohio.
Biochemistry. 1987 Sep 22;26(19):6165-77. doi: 10.1021/bi00393a033.
A human genomic DNA library was screened for the gene coding for human prothrombin with a cDNA coding for the human protein. Eighty-one positive lambda phage were identified, and three were chosen for further characterization. These three phage hybridized with 5' and/or 3' probes prepared from the prothrombin cDNA. The complete DNA sequence of 21 kilobases of the human prothrombin gene was determined and included a 4.9-kilobase region that was previously sequenced. The gene for human prothrombin contains 14 exons separated by 13 intervening sequences. The exons range in size from 25 to 315 base pairs, while the introns range from 84 to 9447 base pairs. Ninety percent of the gene is composed of intervening sequence. All the intron splice junctions are consistent with sequences found in other eukaryotic genes, except for the presence of GC rather than GT on the 5' end of intervening sequence L. Thirty copies of Alu repetitive DNA and two copies of partial KpnI repeats were identified in clusters within several of the intervening sequences, and these repeats represent 40% of the DNA sequence of the gene. The size, distribution, and sequence homology of the introns within the gene were then compared to those of the genes for the other vitamin K dependent proteins and several other serine proteases.
用人凝血酶原的cDNA对人基因组DNA文库进行筛选，以寻找编码人凝血酶原的基因。鉴定出81个阳性λ噬菌体，从中选择3个进行进一步鉴定。这3个噬菌体与从凝血酶原cDNA制备的5'和/或3'探针杂交。测定了人凝血酶原基因21千碱基的完整DNA序列，其中包括一个先前已测序的4.9千碱基区域。人凝血酶原基因包含14个外显子，被13个间隔序列隔开。外显子大小从25到315个碱基对不等，而内含子大小从84到9447个碱基对不等。该基因的90%由间隔序列组成。除了间隔序列L的5'端存在GC而非GT外，所有内含子剪接位点均与其他真核基因中的序列一致。在几个间隔序列中发现了30个Alu重复DNA拷贝和2个部分KpnI重复拷贝，这些重复序列占该基因DNA序列的40%。然后将该基因内含子的大小、分布和序列同源性与其他维生素K依赖性蛋白及其他几种丝氨酸蛋白酶的基因进行比较。
