lacZ转基因鼠体内角蛋白5及10的分布

Distribution of K5, K10 in transgenic mouse with lac Z reporter gene

目的探讨角蛋白５、１０（Ｋ５、Ｋ１０）在活鼠体内的分布。方法（１）用带有β半乳糖甙酶编码的ＬａｃＺ作为报告基因，分别以牛Ｋ５、Ｋ１０［含酶切位点（ＳａｌⅠ）］为启动子的片段插入ｐＬＧＺ及ｐＬＢＳ２质粒。（２）用微注射法将移植基因片段注入鼠受精卵中，保留聚合酶链反应（ＰＣＲ）阳性者的后代作为实验用鼠。（３）切转基因鼠背、尾等冰冻切片，０１％５溴４氯３吲哚βＤ半乳糖甙（χｇａｌ）溶液复盖，核固红补染。结果在鳞状上皮中，Ｋ５分布于表皮基层及毛囊外壁，Ｋ１０分布于基底上层及毛囊外壁；除鳞状上皮外，Ｋ５、Ｋ１０分布于除心脏外的基它脏器中，但脑中无Ｋ５分布。结论ｌａｃＺ转基因鼠有助于基因突变及基因治疗的研究；对Ｋ５、Ｋ１０在体内分布的了解。

Objective To detect the distribution of K5, K10 in live mouse. Methods lacZ gene, which encodes βgalactosidase, as a reporter gene, bovine K5, K10 as prmoter, containing Sal Ⅰ fragment, was inserted into the Sal Ⅰ sites of PLGZ and PLBS2, respectively. The constructs were microinjected into the mice fertilized eggs, and then PCR positive ones were kept. The next generations could be used for experiments. Frozen sections of transgenic mice skin, tail were stained with 0.1% Xgal solution at 37 ℃ overnight and counterstained with nuclear fast red. Result In sguamous epidermis, K5 was distributed in basal and outer root sheath, K10 in suprabasal and inner root sheath. K5 and K10 were also distributed in some other organs except heart. No K5 was seen in the brain. Conclusion lacZ transgenic animal is helpful in the study of gene mutation and treatment. The recognition of K5 and K10 distribution could be a primary step to probe the mechanism of some related diseases and cancers.