贵州农业科学
貴州農業科學
귀주농업과학
GUIZHOU AGRICULTURAL SCIENCES
2014年
6期
6-9
,共4页
水稻%抗虫基因%Cry1C*%表达载体构建
水稻%抗蟲基因%Cry1C*%錶達載體構建
수도%항충기인%Cry1C*%표체재체구건
rice%insect-resistant gene%Cry1C*%construction of expression vector
为水稻害虫生物防治提供菌株，从GenBank中选定Cry 1C 类抗虫基因，将其优化改造后命名为 Cry 1C *，Cry 1C *经 BamH I 和 Spe I 双酶切后与植物表达载体 pTCK303连接，构建 pTCK303-Cry 1C *重组表达载体，将其转化到农杆菌 LBA4404中，同时进行 PCR、双酶切及测序鉴定。结果表明：PCR、双酶切及测序产物与预期扩增片段（2342 bp）相符，重组表达载体 pTCK303-Cry 1C *构建成功；农杆菌LBA4404中含有重组质粒，成功构建了 Cry 1C *农杆菌基因工程菌株。
為水稻害蟲生物防治提供菌株，從GenBank中選定Cry 1C 類抗蟲基因，將其優化改造後命名為 Cry 1C *，Cry 1C *經 BamH I 和 Spe I 雙酶切後與植物錶達載體 pTCK303連接，構建 pTCK303-Cry 1C *重組錶達載體，將其轉化到農桿菌 LBA4404中，同時進行 PCR、雙酶切及測序鑒定。結果錶明：PCR、雙酶切及測序產物與預期擴增片段（2342 bp）相符，重組錶達載體 pTCK303-Cry 1C *構建成功；農桿菌LBA4404中含有重組質粒，成功構建瞭 Cry 1C *農桿菌基因工程菌株。
위수도해충생물방치제공균주，종GenBank중선정Cry 1C 류항충기인，장기우화개조후명명위 Cry 1C *，Cry 1C *경 BamH I 화 Spe I 쌍매절후여식물표체재체 pTCK303련접，구건 pTCK303-Cry 1C *중조표체재체，장기전화도농간균 LBA4404중，동시진행 PCR、쌍매절급측서감정。결과표명：PCR、쌍매절급측서산물여예기확증편단（2342 bp）상부，중조표체재체 pTCK303-Cry 1C *구건성공；농간균LBA4404중함유중조질립，성공구건료 Cry 1C *농간균기인공정균주。
To provide strains for biological control of rice pests,the insect-resistant gene Cry 1C from GenBank was optimized,added BamH I and Spe I restriction enzyme cutting sites at both sides,and artificially synthesized.The modified Bt gene was named Cry 1C * ,which was inserted into pTCK303 after double restriction-enzyme digestion to construct recombinant expression vector pTCK303-Cry 1C * ,and transferred into A.tumefaciens LBA4404.The PCR,double digestion,and sequencing results showed that the recombinant vector was successfully constructed(2 342 bp).The PCR result showed that the expression vector pTCK303-Cry 1C * had been transformed into A.tumefaciens LBA4404,successfully constructing a genetic engineering bacteria.
