摘要
目的 构建重组人脂联素 (ADPN)真核表达质粒 ,为进一步研究ADPN的功能提供实验基础。方法 应用限制性内切酶双酶切载有人脂联素cDNA的重组克隆质粒pMD18 T和真核表达质粒pcDNA 3 1+ ,回收目的基因片段与线性化的真核表达质粒 ,经连接、转化大肠杆菌JM10 9,筛选阳性克隆 ,经酶切和核苷酸序列测定鉴定。结果 重组真核表达质粒经HindⅢ、EcoRⅠ酶切后获得 5 4kb和 80 0bp两个片段 ,大小与理论值一致 ,并经核苷酸序列测定证实。结论 成功地构建了人重组脂联素真核表达质粒。
Objective To provide experimental basis for further investigation on adiponectin (ADPN) function,its eukaryotic recombinant was constructed.Methods The recombinant plasmid pMD18-T/ADPN and eukaryotic expression vector pcDNA 3.1+ were digested by two restrictive endonuclease and ADPN and linear pcDNA 3.1+ were obtained; then, they were linked and translated into JM109; at last, the recombinant pcDNA 3.1+/ADPN was obtained and was identified by digest of restrictive endonuclease and nucleotide sequencing.Results 800 bp fragment and 5.4 kb fragment were consistant with theoretic values after eukaryotic recombinant was digested by HindⅢ and EcoRⅠ.Conclusion the eukaryotic expressing recombinant with complete adiponectin cDNA was constructed
出处
《安徽医科大学学报》
CAS
2004年第3期189-191,共3页
Acta Universitatis Medicinalis Anhui
基金
安徽省卫生厅科研基金资助 (编号 :2 0 0 2A0 0 8)
安徽省省自然科学基金资助 (编号 :0 3 0 43 719)
