Effect of selectable gene to reporter gene ratio on the frequency of co-transformation and co-expression of uidA and hpt transgenes in protoplast-derived plants of tall fescue

被引:11
作者
Bettany, AJE [1 ]
Dalton, SJ [1 ]
Timms, E [1 ]
Dhanoa, MS [1 ]
Morris, P [1 ]
机构
[1] AFRC, Inst Grassland & Environm Res, Aberystwyth SY23 3EB, Ceredigion, Wales
关键词
Festuca arundinacea; fescue; non-selected gene; protoplasts; transformation; transgenic plants;
D O I
10.1023/A:1013894801716
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Forty-six independent transformed plants were regenerated under hygromycin selection from cell-suspension derived protoplasts of Festuca arundinacea (Schreb.) after PEG-mediated transformation. Protoplasts were co-transformed with varying molar gene ratios (0.7:1-6:1) of a marker beta-glucuronidase (uidA) gene and a selective hygromycin (hpt) resistance gene. Logistic regression analysis indicated that, as expected, the proportion of co-transformed plants tended to increase as the proportion of the marker gene was increased. However, although the proportion of plants co-expressing both genes tended to increase up to a molar ratio of 4:1, it appeared to fall at a molar ratio of 6:1. No statistically significant differences were found in the average copy number of the integrated uidA or hpt transgenes, either in GUS expressing, or in non-GUS expressing plants at the different molar ratios. When using naked-DNA gene transformation methods most authors use a molar ratio of 1:1; our data suggest that adding non-selected and selected transgenes at a higher Molar Gene Ratio would probably improve the proportion of plants regenerated which express both transgenes.
引用
收藏
页码:177 / 186
页数:10
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