Dissertation > Biological Sciences > Botany > Plant Cell Genetics > Plant Genetic Engineering

Cloning of CrFtsZ2 Gene from Chlamydomonas Reinhardtii and Expression in E.coli

Author LeiQiYi
Tutor LiuXiangLin
School Capital Normal University
Course Botany
Keywords Chlamydomonas CrFsZ2 The prokaryotic expression Molecular pedigree analysis
CLC Q943.2
Type Master's thesis
Year 2005
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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 .

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