Hewitt J E, Gordon M M, Taggart R T, Mohandas T K, Alpers D H
Gastroenterology Division, Washington University School of Medicine, St. Louis, Missouri 63110.
Genomics. 1991 Jun;10(2):432-40. doi: 10.1016/0888-7543(91)90329-d.
A human gastric intrinsic factor (IF) cDNA clone was isolated using a rat cDNA clone as a probe. Comparison of the predicted amino acid sequence revealed 80% identity of human IF with rat IF. These cDNA clones were used to isolate and map two overlapping clones encoding the human IF gene. The first exon of the cloned region (exon 2) contains 30 bp of the 5' untranslated region, the signal peptide, and the first 8 amino acids of the mature protein. Exons 3-10 encode the remainder of the coding and 3' noncoding regions. Southern analysis of genomic DNA indicated the presence of a single human IF gene and also revealed the presence of strong hybridizing sequences in genomic DNA from monkey, rat, mouse, cow, and human, suggesting that the IF gene is well conserved. The IF gene was localized to human chromosome 11 by concurrent cytogenetic and cDNA probe analysis of a panel of human X mouse somatic cell hybrids. Southern analysis of genomic DNA from patients with congenital pernicious anemia (lacking intrinsic factor) revealed normal restriction fragment patterns, suggesting that a sizable gene deletion was not responsible for the deficiency.
以大鼠cDNA克隆为探针,分离出了人胃内因子(IF)cDNA克隆。对预测的氨基酸序列进行比较后发现,人IF与大鼠IF的同一性为80%。这些cDNA克隆被用于分离和定位两个重叠的克隆,它们编码人IF基因。克隆区域的第一个外显子(外显子2)包含5'非翻译区的30个碱基对、信号肽以及成熟蛋白的前8个氨基酸。外显子3至10编码编码区的其余部分和3'非编码区。基因组DNA的Southern分析表明存在单一的人IF基因,同时还揭示了来自猴、大鼠、小鼠、牛和人的基因组DNA中存在强杂交序列,这表明IF基因高度保守。通过对一组人X小鼠体细胞杂种进行细胞遗传学和cDNA探针同步分析,将IF基因定位到了人类11号染色体上。对先天性恶性贫血(缺乏内因子)患者的基因组DNA进行Southern分析,结果显示限制性片段模式正常,这表明该缺陷并非由大片段基因缺失所致。