小麦抗逆相关基因TaSAP1的5′非翻译区内含子功能分析

常建忠,董春林,张正,乔麟轶,杨睿,蒋丹,张彦琴,杨丽莉,吴佳洁,景蕊莲

Function analysis of 5′ untranslated region introns in drought-resistance gene TaSAP1

Jian-Zhong CHANG,Chun-Lin DONG,Zheng ZHANG,Lin-Yi QIAO,Rui YANG,Dan JIANG,Yan-Qin ZHANG,Li-Li YANG,Jia-Jie WU,Rui-Lian JING

图4 TaSAP1 5UI缺失突变及载体构建 M: marker III (天根); A: 分别用TaspF1/R1 (泳道1)和TaspF2/R2 (泳道2)进行第1轮PCR扩增; B: 用TaspF1/R2进行第2轮PCR扩增, 缺失Intron-1; C: 用TspF1/R3进行PCR扩增获得-2I片段(泳道1); 用TspF1/R3扩增获得Δ2I片段(泳道2); D: 1 ~ 4泳道依次为载体P91z-P1、P91z-(-1I)、P91z-(-2I)和P91z-Δ2I的酶切验证。

Fig. 4 Deletion mutation of TaSAP1 5UIs and construction of expression vectors M: marker III (Tiangen); A: 1st cycle PCR with primers TaspF1/R1 (lane 1) and TaspF2/R2 (lane 2); B: deletion of intron-1 by 2nd cycle PCR with TaspF1/R2; C: -2I and Δ2I fragments were amplified with primers TspF1/R3 (lane 1) and TspF1/R3 (lane 2), respectively; D: lanes 1 to 4 show the double digestion of P91z-P1, P91z-(-1I), P91z-(-2I), and P91z-Δ2I, respectively.