Cloning of CrFtsZ2 Gene from Chlamydomonas Reinhardtii and Expression in E.coli
|School||Capital Normal University|
|Keywords||Chlamydomonas CrFsZ2 The prokaryotic expression Molecular pedigree analysis|
FtsZ is widely present in the function of a protein in prokaryotes and plants , control of prokaryotic cells and plastid division process . Normal E. coli cell division , FtsZ protein earliest gathered into a central division sites in the cell the cyclic skeleton -Z ring to start the process of cell division . Chloroplast is an important organelle in plant cells , and its origins in the early prokaryotes and primitive eukaryotes with photosynthetic capacity endosymbiotic event . Has been confirmed that the endosymbiotic origin of the plant cell chloroplast still FtsZ protein split , but the expression of the protein gene by the chloroplast genome transferred to the nuclear genome . Unlike bacterial cells contain only one FtsZ gene , plant FtsZ genes have evolved to a small family . Previous research suggests that the plant FtsZ protein differences occurred in the early stages of the evolution of terrestrial plants . This thesis, using RT-PCR method the CrFtsZ2 the cDNA was cloned from Chlamydomonas reinhardtii , the full-length cDNA of 1815bp . The gene 10 intron 11 exon , an open reading frame (ORF) 1305bp, encoding 435 amino acids . Sequence homology analysis showed that its encoded protein has the typical characteristics of the FtsZ protein containing the conserved region of the FtsZ protein has a total of two amino acids , as well as similar to tubulin GTP binding sites . Molecular evolution analysis showed that the the Chlamydomonas CrFtsZ1 , CrFstZ2 both the FtsZ1/FtsZ2 subfamily . Further analysis of gene structure , and experimental results show that : endosymbiotic events the original chloroplast genome FtsZ gene immediately transferred to the nucleus , evolved red algae, green algae , in the early stages of differentiation in the green algae , green algae FtsZ gene duplication generated FtsZ1 FtsZ2 . Further identification CrFsZ2 functionality built into the expression vector for prokaryotic expression . The phenotype observed that expression of of exogenous CrftsZ gene significantly affect the normal growth of the host E. coli similar results with prokaryotic FtsZ gene expression , thereby validated conservative of CrFtsZ2 gene structure and function .