Amino acid sequence of the mRNA cap-binding protein from human tissues.
Amino acid sequence of the mRNA cap-binding protein from human tissues.
复制标题
来自人体组织的 mRNA 帽结合蛋白的氨基酸序列。
DOI：
--
复制
发表时间：
1987
影响因子：
11.1
通讯作者：
R. Rhoads
中科院分区：
文献类型：
--
作者：
W. Rychlik;L. Domier;P. R. Gardner;G. M. Hellmann;R. Rhoads
用亲和层析法从人红细胞和兔网织红细胞中纯化了与翻译有关的25 kDa信使核糖核酸结合蛋白(CBP)。测定了8个人和7个兔的胰蛋白酶和V8蛋白水解肽的序列。根据多肽序列数据，合成寡脱氧核苷酸探针，用于筛选人成纤维细胞和淋巴细胞lambda基因文库。从重组Lambda噬菌体插入物中获得的DNA序列被发现编码除一个肽以外的所有肽。根据该DNA序列合成了23个碱基的寡核苷酸，并将其用于启动人胎盘mRNA的合成以构建第三个文库。用22碱基的寡核苷酸筛选，其序列位于23碱基引物的上游，产生了大量的重组噬菌体，大约等于250碱基的插入。从所有噬菌体插入片段编辑的1900个碱基对的cDNA序列似乎代表了CBP(Mr 25,117)的整个一级序列。对人胎盘和HeLa基因的印迹分析显示，CBP基因有多个碱基，从1925到2250个碱基。CBP的氨基酸序列与流感病毒的帽结合蛋白PB2同源。
The 25-kDa mRNA cap-binding protein (CBP) involved in translation was purified by affinity chromatography from human erythrocytes and rabbit reticulocytes. The sequences of eight human and seven rabbit tryptic and V8 proteolytic peptides were determined. Based on the peptide sequence data, oligodeoxynucleotide probes were synthesized and used to screen human fibroblast and lymphocyte lambda cDNA libraries. The DNA sequence obtained from recombinant lambda phage inserts was found to code for all but one peptide. A 23-base oligonucleotide was synthesized based on the DNA sequence and used to prime synthesis of cDNA from human placental mRNA to construct a third library in lambda gt10. Screening with a 22-base oligonucleotide, whose sequence was upstream from the 23-base primer, yielded numerous recombinant phages with approximately equal to 250-base inserts. The 1900-base-pair cDNA sequence compiled from all phage inserts appeared to represent the entire primary sequence of CBP (Mr 25,117). Blot analysis of human placental and HeLa mRNA revealed multiple CBP mRNA species ranging from 1925 to 2250 bases. The amino acid sequence of CBP showed homology to the cap-binding PB2 protein of influenza virus.