Construction of Maize Chloroplast Expression Vector and Transient Expression in Maize Chloroplasts by Detecting GFP

doi:10.7668/hbnxb.2008.04.017

Abstract

Abstract: Tps1 encodes a key enzyme in trehalose synthesis1Maize chloroplast expression vector harboring the yeast Tps1 is constructed in this paper1Firstly, the homologous fragment 16S/ trnI2trnA/ 23S was amplified frommaize chloro2 plast genome by PCR for site2specific integration. Secondly, the yeast Tps1 cassette ( Prrn2 Tps12 Tpsb A2ter ), phos2 phinothricin2resistant gene Bar cassette( Rpsb A pro2 Bar2Rpsb Ater) and Npt / G fp cassette consistingof kanamycin2resistant gene Npt and green fluorescent protein gene G fp ( Prrn2 Npt/ G fp2 RrbcLter) were constructed respectively1 Finally, the three expression cassettes were cloned into the homologous fragment to obtain chloroplast stable expression vector mTKGB.Then mTKGBwas introduced into young leaves of maize through particle bombardment1Two days later, strong green fluorescence was observed in the chloroplasts of some bombarded leaves under confocal laser scanning microscope, indicating that m TKGB could be expressed in maize chloroplasts1The results of this paper provide a strong foundation for genetic engineering drought2resistant maize through chloroplast transformation in the future.

Key words: Maize, Chloroplast expression vector, Transient expression, GFP

CLC Number:

Q786

ZHAO Tong, HUANG Cong-lin, ZHANG Xiu-hai, YU Rong, WANG Yong-qin, WU Zhong-yi. Construction of Maize Chloroplast Expression Vector and Transient Expression in Maize Chloroplasts by Detecting GFP[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2008, 23(4): 72-76. doi: 10.7668/hbnxb.2008.04.017.

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