小叶杨Δ~1-吡咯琳-5-羧酸合成酶(P5CS)基因克隆及在杂种落叶松中的转化

Transformation of Hybrid Larch(Larix Leptolepis x L.Olgensis) with P5CS Gene Cloned from Populus Simonii cDNA Library

以小叶杨为材料构建了干旱胁迫和正常生长条件下的cDNA文库,以特异性引物从中扩增出一条1 850bp大小的DNA片段,经序列分析证实该片段编码Δ1-吡咯琳-5-羧酸合成酶(P5CS)。将该片段构建入植物表达载体pBI121中,在落叶松杂交育种中利用花粉管通道法将带有该P5CS基因的植物表达质粒转化入杂种落叶松,收获球果取出种子,提取转化种子发芽长出幼芽的DNA,特异性PCR扩增和Southern,Western Blotting检测证实落叶松中已导入P5CS基因。

cDNA library were constructed from the drought-treated and normal Populus simonii materials. By using PCR, with specific primers, a segment of 1 500bp was gained and identified as the gene encoding delta 1-pyrroline-5- carboxyhte synthetase P5CS. This segment was constructed into the plant expression vector pBI121. Then the constructed plasmid was transformed into hybrid larch （Larix leptolepis x L. olgensis） by using the Pollen Tube Pathway . The PCR amplification , Southern and Western Blotting hybridization methods and sequence analysis were used to identified the P5CS gene in the transgenic hybrid larch （Larix leptolepis x L. olgensis）.