Chung D W, Que B G, Rixon M W, Mace M, Davie E W
Biochemistry. 1983 Jun 21;22(13):3244-50. doi: 10.1021/bi00282a032.
A total of 148 cDNAs coding for the beta chain of human fibrinogen have been identified from a human liver cDNA library employing a bovine cDNA as a probe. The largest cDNA insert contained 1932 base pairs cloned into the PstI site of plasmid pBR322. This cDNA insert contained 66 base pairs coding for a portion or all of a signal sequence, 1383 base pairs coding for 461 amino acids in the mature protein, a stop codon of TAG, a noncoding region of 431 base pairs, and a poly(A) tail of 19 base pairs. Most of the cDNA inserts coding for the beta chain were found to have a noncoding region of 98 or 167 base pairs rather than 431 base pairs at the 3'-end. The bovine cDNA for the beta chain was also employed as a probe for screening a lambda phage library containing human genomic DNA. Seven positive phage were identified. One of the phage, which contained the entire gene for the beta chain of fibrinogen, was examined by electron microscopy, and portions of its DNA sequence are presented. Seven intervening sequences were identified in the gene for the beta chain of human fibrinogen. The largest intervening sequence (approximately 1.3 kilobases) was found at the 5'-end of the gene and was located between amino acid residues 8 and 9, which are present in fibrinopeptide B. A sequence analysis of the 5'-end of the gene also indicated that the B chain of human fibrinogen contained a signal sequence of either 16, 27, or 30 amino acid residues.
以牛的互补脱氧核糖核酸(cDNA)为探针,从人肝cDNA文库中总共鉴定出148个编码人纤维蛋白原β链的cDNA。最大的cDNA插入片段含有1932个碱基对,克隆到质粒pBR322的PstI位点。该cDNA插入片段包含66个碱基对,编码部分或全部信号序列;1383个碱基对,编码成熟蛋白中的461个氨基酸;一个TAG终止密码子;一个431个碱基对的非编码区;以及一个19个碱基对的聚腺苷酸(poly(A))尾。发现大多数编码β链的cDNA插入片段在3'端具有98或167个碱基对的非编码区,而不是431个碱基对。牛纤维蛋白原β链的cDNA也被用作探针,筛选包含人基因组DNA的λ噬菌体文库。鉴定出七个阳性噬菌体。其中一个含有纤维蛋白原β链完整基因的噬菌体通过电子显微镜检查,并给出了其部分DNA序列。在人纤维蛋白原β链基因中鉴定出七个间隔序列。最大的间隔序列(约1.3千碱基)位于基因的5'端,位于纤维蛋白肽B中存在的第8和第9个氨基酸残基之间。对该基因5'端的序列分析还表明,人纤维蛋白原的B链含有16、27或30个氨基酸残基的信号序列。