Chung D W, Chan W Y, Davie E W
Biochemistry. 1983 Jun 21;22(13):3250-6. doi: 10.1021/bi00282a033.
A number of cDNAs coding for the gamma chain of human fibrinogen have been isolated from a liver cDNA library by employing a synthetic nucleotide mixture as a probe. One of the positive clones was then employed to screen the entire cDNA library of 18000 recombinants, yielding 320 positive clones for the gamma chain. The largest cDNA was 1638 base pairs in length and contained 10 base pairs of poly(G) at the 5'-end followed by 71 base pairs of noncoding nucleotides. The next 78 base pairs coded for a leader sequence that was 26 amino acids in length and included a methionine start signal and a typical hydrophobic core. The following 1233 base pairs coded for 411 amino acids that are present in the mature protein followed by a stop codon of TAA, 207 base pairs of noncoding nucleotides, a poly(A) track of 15 base pairs, and 22 base pairs of poly(C). Specific regions of the cDNA of the gamma chain were then compared with the cDNAs for the alpha and beta chains of human fibrinogen.
通过使用合成核苷酸混合物作为探针,已从肝脏cDNA文库中分离出许多编码人纤维蛋白原γ链的cDNA。然后用其中一个阳性克隆筛选了包含18000个重组体的整个cDNA文库,得到了320个γ链阳性克隆。最大的cDNA长度为1638个碱基对,在5'端含有10个聚(G)碱基对,随后是71个非编码核苷酸碱基对。接下来的78个碱基对编码一个长度为26个氨基酸的前导序列,包括一个甲硫氨酸起始信号和一个典型的疏水核心。随后的1233个碱基对编码成熟蛋白中存在的411个氨基酸,接着是TAA终止密码子、207个非编码核苷酸碱基对、15个碱基对的聚(A)序列和22个碱基对的聚(C)。然后将γ链cDNA的特定区域与人纤维蛋白原α链和β链的cDNA进行比较。