Research on the function of a single transmembrane metalloprotease of mouse ADAM33

去整合素和金属蛋白酶（ADAM）家族是一类具有独特结构域的单跨膜蛋白，包括金属蛋白酶、去整合素和EGF样结构域。迄今已确定了34个亚当斯。17个亚当斯在其金属蛋白酶结构域中含有酶活性所需的独特氨基酸序列（HE-X-H-XX-G-XX-HD）。我们从小鼠脑cDNA文库中分离到一个新的小鼠ADAM 33基因，该基因与非洲爪蟾ADAM 13高度相似，可能在早期神经嵴和神经管的形成中起作用。我们研究了它们的核苷酸和氨基酸序列。小鼠ADAM 33氨基酸序列占42%。与非洲爪蟾ADAM 13同源，但两种蛋白在蛋白质结构上均保守其功能结构域。为了进一步阐明小鼠ADAM 33的功能，我们将氯霉素乙酰转移酶（CAT）基因插入外源启动子和小鼠ADAM 33基因之间，构建了转基因小鼠。然后将构建的DNA显微注射到受精卵中。用ELISA方法分析了该基因导入对转基因小鼠转录水平的影响，未获得明显的高转录小鼠。我们重新建立了基因结构改变的转基因小鼠，小鼠ADAM 33基因直接与其启动子连接。

A disintegrin and metalloprotease (ADAM) family of the single transmembrane proteins has a unique domain structure, containing metalloprotease, disintegrin and EGF-like domains. 34 ADAMs have been identified to date. 17 ADAMs contain the unique amino acid sequence (HE-X-H-XX-G-XX-HD) in their metalloprotease domain required for enzymatic activity. We isolated novel mouse ADAM33 gene from the cDNA library in mouse brain, which is highly resembled the Xenopus ADAM13, suggesting play a role of the formation of early neural crest and neural tube. We investigated their nucleotide and amino acid sequence. The mouse ADAM33 amino acid sequence showed 42%. homology with Xenopus ADAM13, but both proteins were conserved their functional domains in protein structures. We also isolated the human ADAM33 gene, and this amino acid sequence showed the highest homology with mouse ADAM33 (70%).To clarify the function of mouse ADAM33, we generated transgenic mice using the DNA fragment containing chloramphenicol acetyltransferase (CAT) gene inserted between exogenous promoter and mouse ADAM33 gene. And then constructed DNA was microinjected into fertilized eggs. The transcriptional effects of this introduction in generated transgenic mice were analyzed with ELISA method, no obvious highly transcriptional mice were obtained. We resume generating the transgenic mice with alterative gene-structure, mouse ADAM33 gene was directly connected with their promoter.