Molecular cloning of human intestinal mucin cDNAs. Sequence analysis and evidence for genetic polymorphism.
Molecular cloning of human intestinal mucin cDNAs. Sequence analysis and evidence for genetic polymorphism.
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发表时间：
1989-04
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通讯作者：
J. Gum;J. Byrd;J. Hicks;N. Toribara;D. Lamport;Y. Kim
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作者：
J. Gum;J. Byrd;J. Hicks;N. Toribara;D. Lamport;Y. Kim
使用针对人结肠癌异种移植粘蛋白的去糖基化蛋白骨架制备的抗血清筛选人小肠λ gt 11 cDNA文库。从该筛选中分离出三种cDNA，命名为SMUC 40-42。发现这些cDNA均含有69个核苷酸的串联重复序列，其编码富含苏氨酸和脯氨酸的蛋白质共有序列PTTTPITTTTTVTPTPTPTGTQT。用这些cDNA之一SMUC 41探测的RNA印迹在大约7，600个碱基处显示出大的多分散性杂交条带。当使用人小肠、结肠和结肠癌poly（A）+ RNA时，条带强度最强。来自人小肠、结肠和结肠癌细胞的poly（A）+ RNA的体外翻译产生162，000-道尔顿的肽，该肽与去糖基化粘蛋白的抗体免疫沉淀。SMUC 41也被用来探测DNA印迹，这表明限制性片段长度多态性在肠粘蛋白基因的存在。这些发现可能是重要的，在评估发现与几种人类疾病相关的异常粘蛋白。
A human small intestine lambda gt11 cDNA library was screened using antisera prepared against the deglycosylated protein backbone of human colon cancer xenograft mucin. Three cDNAs were isolated from this screening, designated SMUC 40-42. These cDNAs were all found to contain tandem repeats of 69 nucleotides which encoded a threonine- and proline-rich protein consensus sequence of PTTTPITTTTTVTPTPTPTGTQT. RNA blots probed with one of these cDNAs, SMUC 41, exhibited large, polydisperse hybridization bands at approximately 7,600 bases. Band intensities were strongest when human small intestine, colon, and colon cancer poly(A)+ RNA was used. In vitro translation of poly(A)+ RNA from human small intestine, colon, and colon cancer cells produced a 162,000-dalton peptide that was immunoprecipitated with antibodies to deglycosylated mucin. SMUC 41 was also used to probe DNA blots, which indicated the presence of restriction fragment length polymorphisms in the intestinal mucin gene. These findings may be important in assessing the abnormal mucins found associated with several human diseases.