Cloning and Construct of Over-expression Vector of IPA1 cDNA from Nipponbare
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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。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.
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Key words:
- rice /
- IPA1 /
- cDNA cloning /
- vector construction
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[1] 谢华安.中国特别是福建的超级稻研究进展[J].中国稻米,2004,(2):7-9. [2] 周开达,马玉清,刘太清,等.杂交水稻亚种间重穗型组合的选育——杂交水稻超高产育种的理论与实践[J].四川农业大学学报,1995,13(4): 403- 407.
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