Construction of Rice Chloroplast-specific Expression Vectors and Detection on Prokaryotic Expression by EGFP

The elements for rice chloroplast-specific expression vectors were cloned.The fragments of TrnI and TrnA were obtained by PCR from rice,with 1 388 bp and 824 bp in length,respectively,by sequence analysis.Three enhanced green fluorescent protein genes(egfp) with different ribosome binding sites(RBS) were amplified by PCR from pEGFP-N2 plasmid as a template.The promoter Prrn and the terminator TpsbA were from tobacco and were cloned by PCR amplification of pLM21.Three rice chloroplast-specific expression vectors,named as pIA-EGFP1、pIA-EGFP2 and pIA-EGFP3,were constructed and a molecular analysis was conducted.The prokaryotic expression of vectors was tested in E.coli.The vector pIA-EGFP3 with RBS from 5′UTR of λ bacteriophage T7 gene 10 expressed the strongest enhanced green fluorescent protein,indicating that pIA-EGFP3 is a suitable vector for rice chloroplast transformation.