摘要
【目的】构建激活标签载体并将其转入甜瓜中,获得甜瓜T-DNA插入突变体,为甜瓜功能基因组学的研究奠定基础。【方法】用PCR法扩增目的 DNA片段,经EcoRI和Sac I顺序酶切,将目的片段连接到p CAMBIA2301载体上,构建含4×35s Enhancer DNA片段的激活标签载体。以甜瓜品种‘伽师’为材料,利用农杆菌介导的转化体系转化甜瓜愈伤组织,获得甜瓜T-DNA插入突变体。【结果】获得4株T0代甜瓜转化幼苗,通过Kan筛选和PCR鉴定甜瓜转化幼苗的DNA中是否含有目的基因,证明其中3株甜瓜转化幼苗为转基因植株。【结论】获得了突变植株,为甜瓜重要性状基因发掘奠定基础。
【Objective】In order to gain the T- DNA insertional mutants in melons and provide references for further research on the functional genetics of melons,the activation tagging carrier was constructed and transformed into melons.【Method】The target DNA was amplified from Saussurea involucrate Kar. Kir by PCR,after EcoR I and Sac I digestion,the target enhancer was sub- cloned to p CAMBIA2301 vector. The activation tagging carrier was constructed. And the vector was transformed into the callus of Jiashi melons by using Agrobacterium tumefaciens. T- DNA insertional mutants were obtained. 【Result】There were three regenerative plants that were transgenic plants identified by Kanamycin（ Kan） screening and PCR.【Conclusion】The T- DNA insertional mutants were obtained,and the transgenic melons are likely to be used for the discovery of melon important trait gene.
出处
《新疆农业科学》
CAS
CSCD
北大核心
2016年第6期1048-1053,共6页
Xinjiang Agricultural Sciences
基金
国家大学生创新创业实训项目
国家自然科学基金项目(3126025)~~
作者简介
许荣华（1994-），女，重庆人，本科，研究方向为植物生理生化与分子生物学，（E—mail）952138547@qq．com
通讯作者（cotrespondjng author）：李冠（1949-），男，湖北人，教授，博士生导师，研究方向为植物生理生化与分子生物学，（E-mail）gLlanli@xju．edu．cn
