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通过粒子轰击导入后玉米中可选择和不可选择转基因的结构与功能

Structure and function of selectable and non-selectable transgenes in maize after introduction by particle bombardment.

作者信息

Register J C, Peterson D J, Bell P J, Bullock W P, Evans I J, Frame B, Greenland A J, Higgs N S, Jepson I, Jiao S

机构信息

ICI Seeds Research Department, Slater, IA 50244.

出版信息

Plant Mol Biol. 1994 Sep;25(6):951-61. doi: 10.1007/BF00014669.

Abstract

Zea mays transformants produced by particle bombardment of embryogenic suspension culture cells of the genotype A188 x B73 and selected on kanamycin or bialaphos were characterized with respect to transgene integration, expression, and inheritance. Selection on bialaphos, mediated by the bar or pat genes, was more efficient than selection on kanamycin, mediated by the nptII gene. Most transformants contained multicopy, single locus, transgene insertion events. A transgene expression cassette was more likely to be rearranged if expression of that gene was not selected for during callus growth. Not all plants regenerated from calli representing single transformation events expressed the transgenes, and a non-selectable gene (uidA) was expressed in fewer plants than was the selectable transgene. Mendelian inheritance of transgenes consistent with transgene insertion at a single locus was observed for approximately two thirds of the transformants assessed. Transgene expression was typically, but not always, predictable in progeny plants--transgene silencing, as well as poor transgene transmission to progeny was observed in some plant lines in which the parent plants had expressed the transgene.

摘要

通过对基因型为A188×B73的胚性悬浮培养细胞进行粒子轰击产生的玉米转化体,在卡那霉素或双丙氨膦上进行筛选,并对其转基因整合、表达和遗传特性进行了表征。由bar或pat基因介导的双丙氨膦筛选比由nptII基因介导的卡那霉素筛选更有效。大多数转化体包含多拷贝、单一位点的转基因插入事件。如果在愈伤组织生长期间未选择该基因的表达,则转基因表达盒更有可能发生重排。并非所有从代表单一转化事件的愈伤组织再生的植株都表达转基因,并且与可选择的转基因相比,非选择基因(uidA)在更少的植株中表达。在所评估的约三分之二的转化体中观察到与单一位点转基因插入一致的转基因孟德尔遗传。转基因表达在子代植株中通常(但并非总是)是可预测的——在一些亲本植株已表达转基因的株系中,观察到转基因沉默以及向子代的转基因传递不佳。

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