文章摘要
胡圳圳,郑大同.Galectin-3重组真核表达载体的构建及其在NIH/3T3细胞中的表达[J].南京医科大学学报,2014,(1):
Galectin-3重组真核表达载体的构建及其在NIH/3T3细胞中的表达
Construction of the recombinant eukaryote expression vector encoding gene of galectin-3 and its expression in NIH/3T3 cells
投稿时间:2013-07-29  修订日期:2013-09-22
DOI:10.7655/NYDXBNS201401035
中文关键词: 半乳糖凝集素-3  NIH/3T3细胞  细胞增殖
英文关键词: Galectin-3  NIH/3T3 cell  Cell proliferation
基金项目:南京医科大学科技发展基金重点项目(2012NJMUZ088)
作者单位E-mail
胡圳圳 南京医科大学第二附属医院 Ansel.zzhu@gmail.com 
郑大同 南京医科大学第二附属医院 dtzheng@gmail.com 
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中文摘要:
      目的:构建半乳糖凝集素-3 (Gal-3) 的真核表达载体,在NIH/3T3细胞中表达,并检测其表达。方法: 通过DNA重组技术和PCR方法从人肿瘤细胞克隆Gal-3基因,插入真核表达载体pEGFP-N1中,通过酶切和测序鉴定重组载体的正确性;采用脂质体转染技术将重组质粒pEGFP-Gal-3瞬时转染NIH/3T3细胞,经荧光和Western blot方法检测Gal-3表达,MTT法检测Gal-3对NIH/3T3细胞增殖的影响。结果: 限制性内切酶鉴定和核酸序列测序证实成功构建含Gal-3的重组真核表达载体pEGFP-Gal-3。以重组质粒瞬时转染NIH/3T3细胞,检测到Gal-3蛋白表达,并证实Gal-3蛋白促进NIH/3T3细胞增殖。结论: 成功构建的pEGFP-Gal-3真核表达载体在小鼠NIH/3T3细胞中成功表达,其促进NIH/3T3细胞增殖。
英文摘要:
      Objective: To construct the recombinant eukaryote expression vector containing Gal-3 gene and detect the protein expression in NIH/3T3 cells. Methods: Gal-3 gene was obtained by PCR from cancer cells. PCR product of Gal-3 was then clone into the eukaryote expression vector pEGFP-N1, then the recombinant vector pEGFP-Gal-3 was transfected into the NIH/3T3 cells. After transfection of the recombinant vector, the protein expression of Gal-3 in NIH/3T3 cells was detected by Western blot. The effect of Gal-3 on cell proliferation was investigated by MTT assay. Results: The recombinant eukaryote expression vector encoding Gal-3 was constructed successfully. The expression of Gal-3 in NIH/3T3 cells could be detected by Western blot. In addition, Gal-3 protein could promote the proliferation of NIH/3T3 cells. Conclusion: The new recombinant expression vector pEGFP-Gal-3 was constructed and expressed successfully in NIH/3T3 cells. Furthermore, our study found that Gal-3 protein could promote the proliferation of NIH/3T3 cells.
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