水稻日本晴<em>IPA1</em> cDNA的克隆及植物过表达载体的构建

水稻日本晴IPA1 cDNA的克隆及植物过表达载体的构建

1.
福建省农业科学院水稻研究所, 福建福州 350019;
2.
农业部华南杂交水稻种质创新与分子育种重点实验室/ 福州国家水稻改良分中心/福建省作物分子育种工程实验室/福建省水稻分子育种重点实验室, 福建福州 350003;
3.
福建省作物种质创新与分子育种省部共建国家重点实验室培育基地, 福建福州 350003;
4.
水稻国家工程实验室, 福建福州 350003;
5.
杂交水稻国家重点实验室华南基地, 福建福州 350003

基金项目:

国家转基因作物新品种培育重大专项(2011ZX08001-004);福建省自然科学基金项目(2012J01109);福建省农业科学院青年基金项目(2011QA-3);福建省财政专项——福建省农业科学院科技创新团队建设项目(STIF-Y04)

1.
Rice Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou, Fujian 350019, China;
2.
Key Laboratory of Germplasm Innovation and Molecular Breeding of Hybrid Rice for South China, Ministry of Agriculture, P.R China/Fuzhou branch, National Rice Improvement Center of China/Fujian Engineering Laboratory of Crop Molecular Breeding/ Fujian Key Laboratory of Rice Molecular Breeding, Fuzhou, Fujian 350003, China;
3.
Incubator of National Key Laboratory of Fujian Germplasm Innovation and Molecular Breeding Between Fujian and Ministry of Sciences & Technology, P.R.China, Fuzhou, Fujian 350003, China;
4.
National Rice Engineering Laboratory of China, Fuzhou, Fujian 350003, China;
5.
South China Bases of National Key Laboratory of Hybrid Rice for China, Fuzhou, Fujian 350003, China

摘要: 采用RT-PCR技术,从水稻日本晴中扩增获得了理想株型基因IPA1(Ideal Plant Architecture,IPA)的cDNA片段。测序结果表明,克隆获得的目的片段长为1 257 bp,包含完整的CDS,共编码 418个氨基酸,预测表明该蛋白分子质量为42.51 kD,理论等电点为9.37。将该片段连接至载体pHI,通过SmaI、SacI酶切鉴定及测序验证,结果表明成功构建了pHI-IPA1植物过表达载体,并将载体质粒转化农杆菌,为进一步进行籼稻理想株型基因的遗传转化奠定基础。
- 水稻 /
- IPA1 /
- cDNA克隆 /
- 载体构建
Abstract: A cDNA encoding IPA1 was isolated from Nipponbare by RT-PCR, sequencing results showed that the sequence consists of 1 257 bp with a complete CDS and encodes a protein of 418 amino acids. In addition, the prediction results indicated that molecular weight of target protein was 42.51 kD and theoretical isoelectric point was 9.37. The plant expression vector was constructed by ligating the cDNA fragment into expression vector pHI, then the recombinant plasmid pHI-IPA1 was confirmed by restriction enzyme digestion analysis with SmaI、SacI, and further verified by DNA sequencing. Vector plasmid was transformed into Agrobacterium tumefaciens, which will lay foundation for rice genetic transformation.
- rice /
- IPA1 /
- cDNA cloning /
- vector construction

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