以木糖异构酶基因为筛选标记的玉米遗传转化  

Maize Transformation Using Xylose Isomerase Gene as a Selection Marker

作　　者：郭新梅[1,2] 张晓东[1] 梁荣奇[1] 张立全[1] 陈耀锋[3] 

机构地区：[1]北京市农林科学院农业生物技术研究中心 [2]青岛农业大学生命科学学院,青岛266109 [3]西北农林科技大学农学院

出　　处：《植物生理与分子生物学学报》2007年第6期547-552,共6页Journal Of Plant Physiology and Molecular Biology

基　　金：国家自然科学基金项目(No.30571170)资助。

摘　　要：利用木糖异构酶基因作为筛选标记可以在含有不同浓度木糖的培养基上筛选出玉米再生植株,其中50%～100%木糖浓度的总体筛选效果较好,但不同玉米基因型之间筛选的最佳浓度差异很大。通过DNA点杂交、PCR及PCR-Southern印记法检测表明,木糖异构酶基因已经整合到转基因植株中。以木糖作为筛选剂,可以减小潜在的生物安全隐患。The xylA gene, encoding xylose isomerase, was cloned as a 1 342-bp BamHI/SacI fragment from the E. coli. As a selection marker, the xylA gene was fused between the enhanced CaMV 35S promoter （E35S） and terminator （35St） in pBAC413 （Fig.2）. pBAC413 was constructed to prevent the expression of sbelIb in maize. PDS 1000/He was used to bombard maize calli, which were induced to form by the elite inbred lines. The selection was carried out on the media containing concentrations of xylose from 0 to 100%. The results showed that the media containing 50% to 100% D-xylose were better, but differed with the genotype of maize （Tables 1 and 2）.Successful integration of xylA gene into the maize genome was confirmed by DNA dot blotting, PCR and PCR-Southern hybridization （Figs.4 to 6）. A method was established in which transformed maize cells were successively screened on a medium containing xylose instead of antibiotic and herbicide for bio-safety.

关 键 词：木糖 木糖异构酶 筛选标记基因 玉米 

分 类 号：S513[农业科学—作物学] Q943.2[生物学—植物学]