嵌合基因δKρF在转基因小鼠内的组织特异性表达  

Tissue-specific Expression of the Chimeric Gene δKpF in Transgenic Mice

作　　者：胡以平[1] 秦世真 许月芳[1] 刘祖洞[1] 

机构地区：[1]复旦大学遗传学研究所,上海200433

出　　处：《Acta Genetica Sinica》1992年第1期27-33,共7页

基　　金：中国科学院科学基金~~

摘　　要：本文报告了鸡δ晶体蛋白基因的启动子,被Friend脾病灶形成病毒(SFFV)的长末端重复顺序(LTR)取代后的嵌合基因δKpF在转基因小鼠内的表达特性。采用显微注射方法,将含有δKpF基因的重组质粒pδKpF DNA导入小鼠受精卵的雄原核,再将其植入假孕小鼠的输卵管内,经生长发育,产出小鼠35只。以DNA-DNA分子杂交分析,发现其中1只小鼠的基因组DNA中整合有所导入的δKpE基因。再以免疫酶标记方法检测,发现δKpF基因在转基因小鼠内能够表达,而且具有组织特异性。作者认为,与δ晶体蛋白基因组织特异性表达有关的调节顺序位于+677bp之后。这与Hayashi等(1985)以培养细胞研究的结论不一致。The expression of the chimeric gene δKpF in which the promoter region of δ-crystallin gene was replaced with the long terminal repeats (LTR) of Friend spleen focus forming virus (SFFV) was studied in transgenic mice for approaching the regulatory mechanism of the expression of chicken δ-crystallin gene.The recombinant plasmid (pδKpF) DNA containing δKpF gene was microinjected into male pronuclei of fertillized mouse eggs. The injected eggs were implanted into the oviducts of pseudopregnant female mice and allowed to develop to term. Subsequently, 35 mice from those egges were born. The mice were analyzed at 2 weeks of age with respect to gene integration and expression by the method of DNA blot hybridization and immunoenzymologic assay. DNA-DNA hybridization indicatel that the genomic DNA from one of the mice retained the sequences that hybridized strongly with the probe, pδKpF. Enzyme linked immunoso-rbent assay (ELISA) showed that δKpF gene was expressed in the lens, but not in the brain, muscle and liver.These results demonstrate that the essential regulatory elements for tissue-specific expression of chicken δ-crystallin gene reside in the sequences downstream from + 677hp.

关 键 词：嵌合基因 转基因小鼠 组织特异性 

分 类 号：Q75[生物学—分子生物学]