CAJ | 학술논문

根据植物基因的结构特征,合成了CrylAc活性杀虫蛋白的编码序列并与内质网定位肽编码序列组成嵌合杀虫蛋白基因Bt29K.构建了含Bt29K基因及慈菇蛋白酶抑制剂B(API-B)基因表达框的双抗虫基因植物表达载体.通过根癌土壤杆菌(Agrobacteriumtumefaciens(Smith et Townsend)Conn LBA4404)介导转化了棉花(Gossypium hirsu-tun L.)的两个生产品种(系).根据抗棉铃虫(Heliothis armigera)试验及农艺性状的观察调查结果,经6代筛选,获得了抗棉铃虫90.0%～99.7%且农艺性状优良的9个双价抗虫棉纯合品系.分子生物学分析结果表明,两个抗虫基因在棉花基因组中的插入拷贝数为1个或2个.活性Cry1Ac和API-B蛋白在转基因抗虫棉株系中的表达量分别约占总可溶性蛋白的0.17%和0.09%.对双抗纯合系植株及仅转Bt基因的棉花纯合系抗虫性检测结果表明前者的抗虫性明显高于后者,因此推断本研究采用的双抗虫基因表达载体构建策略是合理的.
근거식물기인적결구특정,합성료CrylAc활성살충단백적편마서렬병여내질망정위태편마서렬조성감합살충단백기인Bt29K.구건료함Bt29K기인급자고단백매억제제B(API-B)기인표체광적쌍항충기인식물표체재체.통과근암토양간균(Agrobacteriumtumefaciens(Smith et Townsend)Conn LBA4404)개도전화료면화(Gossypium hirsu-tun L.)적량개생산품충(계).근거항면령충(Heliothis armigera)시험급농예성상적관찰조사결과,경6대사선,획득료항면령충90.0%～99.7%차농예성상우량적9개쌍개항충면순합품계.분자생물학분석결과표명,량개항충기인재면화기인조중적삽입고패수위1개혹2개.활성Cry1Ac화API-B단백재전기인항충면주계중적표체량분별약점총가용성단백적0.17%화0.09%.대쌍항순합계식주급부전Bt기인적면화순합계항충성검측결과표명전자적항충성명현고우후자,인차추단본연구채용적쌍항충기인표체재체구건책략시합리적.
A chimeric gene, Bt29K, composed of coding sequences of activated Cry1Ac insecticidal protein and an endoplasm reticulum-retarding signal peptide, was synthesized. A plant expression vector containing two expression cassettes for the Bt29K and API-B genes was constructed. These two insect-resistant genes were transferred into two cotton ( Gossypium hirsutum L.) varieties (or lines) via Agrobacterium-mediated transformation and nine homozygous transgenic cotton lines showing a mortality of 90.0% -99.7% to cotton ballworm ( Heliothis armigera ) larvae and good agronomic traits were selected through six generations. Molecular biology analysis revealed that one or two copies of the insecticidal protein genes were integrated into the transgenic cotton genome and activated Cry1Ac and API-B protein expression was at a level of 0.17% and 0.09% of the total soluble protein in the transgenic cotton leaves, respectively. Comparison of the insect-resistance of the homozygous lines expressing the activated chimeric Cry1Ac and API-B with that expressing Cry1Ac only revealed that the insect-resistance of the former is apparently higher than the latter. These results also indicate that the strategy to construct a plant expression vector expressing two different insect-resistant genes reported here is reasonable.