Cloning and expression of the murine gene and chromosomal location of the human gene encoding N-acetylglucosaminyltransferase I.
Cloning and expression of the murine gene and chromosomal location of the human gene encoding N-acetylglucosaminyltransferase I.
复制标题
编码 N-乙酰氨基葡萄糖转移酶 I 的小鼠基因的克隆和表达以及人类基因的染色体定位。
DOI：
10.1093/glycob/2.4.383
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发表时间：
1992
期刊：
影响因子：
4.3
通讯作者：
Stanley,P
中科院分区：
文献类型：
--
作者：
Kumar,R;Yang,J;Eddy,RL;Byers,MG;Shows,TB;Stanley,P
从F9畸胎癌细胞文库中分离的小鼠c DNA克隆[Kumar，R.，Yang，J.，Larsen，R.D.和Stanley，P.(1990)Proc.钉子。阿卡德科学公司。USA，87,9948-9952]已测序。核苷酸和推导的氨基酸序列与先前描述的人和兔的GlcNAc-TI cDNAs高度同源。小鼠除前34个氨基酸外的1250个氨基酸的编码区在大肠杆菌中得到了诱导表达，并产生了一个显著的摩尔融合蛋白。WT˜45 kDa的存在与细胞裂解物中高水平的GIcNAc TI活性有关。从该cDNA产生的探针被用来证明GlcNAc-TI基因在哺乳动物中以单拷贝形式存在，并且在酵母、海胆、果蝇和线虫的DNA中(在低严格杂交条件下)没有检测到同源基因。从小鼠肝脏文库中分离出与GlcNAc-TI cDNA产生的探针杂交的基因组DNA克隆。亚克隆基因组DNA片段的限制性内切酶分析、Southern杂交和DNA序列分析以及聚合酶链式反应(PCR)产物证明，该基因的编码区和3‘非翻译区位于单个外显子中。然而，小鼠GlcNAc-TI基因(mGat-1)至少包括编码区的一个额外外显子5‘。人-鼠杂交体细胞DNA的Southern分析和原位杂交定位了人GlcNAc-TI基因(MGAT-1)位于5号染色体q31.2和q31.3之间，该区域与小鼠11号染色体的一个区域同源。对成年小鼠组织的Northern分析显示，两个GlcNAc-TI基因转录本在不同组织中差异表达。
A mouse cDNA clone previously isolated from an F9 teratocarcinoma cell library and shown to conferN-acetylglucosaminyltransferase I (GlcNAc-TI) activity on Lec1 Chinese hamster ovary (CHO) cell transfectants [Kumar, R., Yang,J., Larsen,R.D. and Stanley,P. (1990)Proc. Nail. Acad Sci. USA, 87, 9948-9952] has been sequenced. The nucleotide and deduced amino acid sequences are highly homologous to previously described human and rabbit GlcNAc-TI cDNAs. A 1250 bp portion of the mouse cDNA encoding all but the first 34 amino acids of the deduced protein sequence was inducibly expressed inEscherichia coliandgave rise to a prominent fusion protein of mol. wt ˜45 kDa whose presence correlated with high levels of GIcNAc TI activity in cell lysates. Probes generated from the cDNA were used to show that the GlcNAc-TI gene is present in a single copy in mammals and that a homologous gene was not detectable (under low-stringency hybridization conditions) in DNA from yeast, sea urchin,DrosophilaorChaenorhabditis elegans. Genomic DNA clones that hybridized to probes generated from the GlcNAc-TI cDNA were isolated from a mouse liver library. Restriction analyses, Southern hybridization and DNA sequence analyses of subcloned genomic DNA fragments and a polymerase chain reaction (PCR) product provided evidence that the coding and 3′ untranslated regions of the cDNA reside in a single exon. However, the mouse GlcNAc-TI gene (Mgat-1) includes at least one additional exon 5′ of the coding region. Southern analyses of DNA from mouse-human somatic cell hybrids andin situhybridization were used to locate the human GlcNAc-TI gene (MGAT-1) between positions q31.2 and q31.3 on chromosome 5, a region of chromosome 5 that is syntenic with a region of mouse chromosome 11. Northern analyses of adult mouse tissues revealed two GlcNAc-TI gene transcripts that are differentially expressed in different tissues.