The Gene Cloning And The Prediction, Comparison And Evolutionary Analysis Of The Structure Of Chlamydomonas Reinhardtii CrFtsZ1 And Oryza Sativa OsFtsZ1

As fomentation of temperature-sensitive protein, FtsZ is a normal protein in prokaryotic. As a kind of GTPas, FtsZ can intergrate to filamentation or tube when depending-GTP. In fact it is a 400aa bacteria conservative protein whose structure and function similar to that of tubulin's. FtsZ also be a key cytoplasm moving cell division protein. On the cell membrane division site where it forms a ring-like structure which acts as a framework and disassembled when cell division finished. Surprisingly, in both of archaebacteria and chloroplast, FtsZ is the main component in the process of division but it isn't effective in mitochodria division, instead is a dynamin protein. Considering this interesting character of FtsZ, the conservation, the differentiation of the two families and the evolution steps attract us and make us carried on the following study.Formers has done enough fine reaserch works on prokaryotic FtsZ, in this paper we try to find the evolutionary clew of its transfering from junior algae to senior plant by choosing the single-cell chlorophyta -Chlamydomonas reinhardtii and gramineae poaceae - Oryza sativa as the experiment material.We cloned OsFtsZI reported in rice by RT-PCR and the CrFtsZI in chlamydomonas by bioinformatic technique and lab works. At first we got a fragment of CrFtsZI by a pair of primer divised according to FtsZ conservative region. Then we sequencing it and blasting in dbEST bank, alignment and splicing the overlaped EST group. At last the completed new gene CrFtsZI was producted. In order to check if it is the aim gene, we devised PCR with a new pair of primer, sequenced and registered the product with registration number:AF449446. Moreover we forecast and analysis the primary, secondary, tertiary and quaternary structure of the three protein: OsFtsZI, CrFtsZI and CrFtsZ2 which has already cloned by our team before. After that we construct FtsZ molecular evolution tree to site them in.Research shows that both of OsFtsZI and CrFtsZI can be translated by frame2. Alpha helix, extended strand and random coil are the major structures of all three protein; The sites where special coils appeared are conservative and more complex in FtsZ1 than in FtsZ2, and more intricate situation in rice than in chlamydomonas. FtsZ is not transmembrane protein, but much more obviously difference of anchor sites on membrane in the two families than in varied species. Otherwise leading peptide exists in FtsZ1 family only. Although it has predicted that different FtsZ families share the same tertinary structure but they also similar to that of tubulin's. In the quaternary structure: tubulin domain, FtsZ1/2 conservative domain and low-complexity domain are the necessary domains in FtsZ, CrFtsZ containing Chlamydomonas special low-complexity domain--Ala rich region confers that more low-complexity domains exited in chlamydomonas. In researching, we found that there is a special coil sequence near the N-end of CrFtsZI whose function we guess should responses for the special character of chloroplast in chlamydomonas perhaps.The evolution analysis suggests that FtsZ must has been experienced an evolutionary process, until in modern bacteria world it has differentiated into two families. That between varied bacteria other than between different families there lies much difference is contrary to that in plant world. However bacteria FtsZ has some cognation relationship with tubulin, perhaps they have the same ancestor.