Cloning of Coat Protein Gene of Tomato Yellow Leaf Curl Virus Strain from Shouguang Region of Shandong Province and Its Expression in E. coli BL21(DE3)
1Institute of Vegetables and Flowers,Weifang College of Science and Technology,Shouguang 262700,Shandong,China|2College of Life Science,Qingdao Agricultural University,Qingdao 266109,Shandong,China|3College of Plant Protection,Shandong Agricultural University,Tai’an 271018,Shandong,China
Abstract:The PCR test was carried out on tomato(Lycopersicon esculentum Mill.)leaves which were infected by TYLCV in the greenhouse in Shouguang region of Shandong Province using the prime PA/PB.The results showed the yellow leaf curl disease found in Shouguang was caused by TYLCV.A DNA fragment(770 bp in length)encoding TYLCV coat protein(TYLCV-CP)was amplified and inserted into pEASY-T1 Simple vector.After double digestion with Xho I and EcoR I,the fragment was inserted into vector pET-32a.A 50 kD recombinant protein of TYLCV-CP from isolate of Shouguang highly expressed by IPTG in E. coli BL21(DE3)showed a high antigenic activity.
乔宁,李美芹,郭宝太,刘永光,裴华丽,竺晓平. 山东寿光地区番茄黄化曲叶病毒外壳蛋白基因克隆及其在大肠杆菌中的表达[J]. 中国蔬菜, 2012, 1(6): 35-40.
QIAO Ning1,LI Mei-qin1*,GUO Bao-tai2,LIU Yong-guang1,PEI Hua-li1,ZHU Xiao-ping3. Cloning of Coat Protein Gene of Tomato Yellow Leaf Curl Virus Strain from Shouguang Region of Shandong Province and Its Expression in E. coli BL21(DE3). , 2012, 1(6): 35-40.