黑素瘤抑制蛋白启动子在小鼠体内的表达活性

Investigation on the expression of melanoma inhibitory protein promoter in vivo

目的：观察黑素瘤抑制蛋白（ＭＩＡ／ＣＤ－ＲＡＰ）启动子在小鼠体内的表达活性。方法：利用ＰＣＲ技术扩增２．２ ｋｂ 的ＭＩＡ／ＣＤ－ＲＡＰ启动子，分子克隆技术构建ＭＩＡ 启动子－半乳糖苷酶（２．２ｌａｃＺ）载体，显微注射法将纯化的２．２ｌａｃＺＤＮＡ 注入来自Ｂ６ＳＪＬ杂交小鼠受精卵的原核以制备转基因小鼠。提取幼鼠尾ＤＮＡ，用ｌａｃＺ特异性引物筛选阳性转基因小鼠。结果：８只首建小鼠染色体基因组整合有２．２ｌａｃＺ融合基因。Ｘ－ｇａｌ全胚胎染色发现其中３ 只转基因小鼠在软骨和乳腺组织中均可见转基因的表达。结论：２．２ ｋｂ

Objective: To investigate the expression pattern of 2.2 kb melanoma inhibitory protein (MIA) promoter in vivo . Methods: 2.2 kb of MIA promoter was amplified by PCR and cloned into lacZ expression vector placF by recombinant techniques to generate MIA promoter lacZ construct (2.2lacZ). Purified DNA was microinjected into the pronuclei of fertilized eggs from B6SJL hybrid to produce transgenic mice. Founder mice were identified by PCR assays of the genomic DNA extracted from tails. Results: Eight transgenic founder mice carrying 2.2lacZ fusion gene were obtained. Whole mount staining with X gal demonstrated 2.2lacZ generated expression in cartilage and mammary glands in 3 transgenic mice. Conclusion: The 2.2 kb of the MIA promoter contains cis acting elements which can generate tissue specific expression in vivo .