拟南芥AtMYB61基因的克隆和表达载体构建.pdf

第34 卷第5 期 合肥工业大学学报( 自然 科 学 版 ) V ol. 34 N o . 5 2011 年5 月 JO U RN AL O F H E FEI U N IV E RSIT Y OF T E CH N OL O GY M ay 2011 Doi 10 . 3969/ j . is sn . 100 35060. 2011. 05. 027 A tM YB 6 1 王 云, 任永兵, 杨 硕, 韩 宇, 陶 杨, 曹树青 ( , 230009) RN A , R TPCR A tM YB 6 1 cDN A , p U CmT , PCR cDN A A tM YB 61 At M YB61pU CmT , Bst E Bg l , p CA M BIA2 30 1 PCR p CAM BIA 2301 At M YB61, A tM YB 61 , A tM YB 61 : ; A tM YB 61 ; Q 37 ; Q 782 A 100 35060( 20 11) 0507 5705 Cl oning of Arabidopsis thaliana A tM YB 61 gene and the expression vector construction WA N G Y un , R EN Y ongbing, Y A N G Shuo, H A N Y u, T A O Y ang , CA O Shuqing ( Sch ool of Biot echnology and F ood En gi neering , Hefei U niv ersit y of T echnology , H ef ei 230009, Ch ina) Abstract: T o tal RN A w as ext ract ed fro m A rabidop sis t haliana seedling s and u sed as t he t em plat e t o am plif y t he f ull leng th of cDN A o f A tM YB 61 gene by RTP CR technolog y, and t he g ene f ragm ent w as subsequent ly cloned int o plant pU CmT v ect or . T he result s o f b act erial co lony P CR , enzym e analysis and cDN A sequ encing confirm ed t h at t he A rabidopsis t h aliana A tM YB 61 g ene w as su cces sf ully cloned. A tM YB 61 gene w as cut com plet ely fr om A t M YB61p U CmT v ecto r by Bst E and Bgl , and the g ene frag ment w as clo ned int o plant ex pres sion v ect or p CA MBIA 2301. T he results o f bacterial colo ny PCR and enzyme analy sis show ed t he su ccessful co nst ruct io n o f plant expressio n vecto r pCA M BIA 2301A t M YB61. In addit io n, the recom binant expressio n v ect or w as carried int o A gro bact erium tumef acien s by elect rot ran sf ormat ion , and th