中文摘要：

目的构建β-catenin真核表达载体并证实融合蛋白在细胞内的表达及定位。方法提取工具细胞NIH3T3的mRNA，反转录为cDNA。PCR扩增β-catenin基因cDNA全长，并将其亚克隆至pEGFP-C1表达载体中。进一步将构建的重组质粒进行酶切和测序鉴定，并转染到工具细胞NIH3T3细胞中，提取细胞蛋白进行Westemblot检测。最后利用激光扫描共聚焦显微镜观察pEGFP-β-catenin在NIH3T3细胞内的定位。结果β-catenin基因cDNA全长克隆到了真核表达载体pEGFP-C1中，酶切鉴定片段为2346bp，并测序成功。Westernb10t检测到了GFP-β-catenin融合蛋白表达，分子量约为115kDa。pEGFP-β-catenin在工具细胞NIH3T3细胞中主要定位于细胞膜和细胞质。结论成功构建了β-catenin基因cDNA全长的真核表达载体，pEGFP-β-catenin蛋白在NIH3T3细胞中主要定位于细胞膜和细胞质。

英文摘要：

Objective To construct the expression plasmid of beta-catenin （or β -catenin） gene and identify the expression and localization of its fusion protein. Methods Total mRNA was extracted from NIH3T3 ceils, cDNA was formed by reverse transcription. The β-catenin coding sequence was amplified by polymerase chain reaction （PCR） and subcloned into pEGFP-C1 vector. After the target region was identified by enzyme digestion and sequencing, the plasmid was transfected into NIH3T3 cells. The expression of the recombinant plasmid in NIH3T3 cells was detected by Western blot. The localization of pEGFP-β-catenin in NIH3T3 cells was observed with laser scanning confocal microscopy. Results The length of the fragment identified by restriction enzyme digestion was 2346bp. The expression of pEGFP- β - catenin fusion protein with a molecular weight of 115kDa was detected by Western blot. The pEGFP- β -catenin fusion protein was mostly localized in the membrane and cytoplasm of NIH3T3 cells. Conclusion The recombinant plasmid of β -catenin gene was successfully cloned into eukaryotic expressing vector, and the pEGFP-β-catenin fusion protein was mostly localized in the membrane and cytoplasm of NIH3T3 ceils.