Study On The Relationship Between Cell Division Of Anabaena PCC7120 And The Expression Of A115278 Gene

CDC2(CDKI,cyclin-dependent protein kinase 1),a serine/threonine protein kin-ase,is an important protein kinase during cell division cycle in eukaryotic cells.Hom-ologous genes have been found in almost all eukaryotic genomes,CDC2 is highly co-nserved.It’s known that there are a lot of serine/threonine protein kinases in cyano-bacteria,in this paper,the correlation between cell division with the expression of all-5278,which is homologous gene of CDC2 in Anabaena sp.PCC7120 was studied.1.2.1 ObjectivesTo find out the effects of roscovitine,the inhibitor of serine/threonine protein kin-ases,on the growth and viability of Anabaena sp.PCC 7120,Synechocystis sp.PCC 6803 and Synechococcus sp.PCC 7002.To detect the expression of all5278 gene and i-ts protein when cells divide is exuberant.1.2.2 MethodsBioinformatics analysis:using bioinformatics databases and software from the Int-emet to analysis all5278 protein.Using the CDC2 inhibitor roscovitine examined the relationship between cyan-obacterial cell viability and expression of al15278:divided Anabaena sp.PCC 7120,Synechocystis sp.PCC 6803 and Synechococcus sp.PCC 7002 into different groups r-espectively,treated with different concentration of roscovitine for 4 hours and 4 days.Determine the concentration and cell viability of cells from each group.Induce Anabaena sp.PCC 7120 to enter cell division by using red light,exa-mined the correlation between the expression of all5278 with cell division:culture Anabaena sp.PCC 7120 in the nitrogen-shortage BG-11 medium,and induce it to ent-er cell division by using red light.After treated for Oh,1h,2h,3h,4h,5h,6h,7h,8h,9h,10h,11h,12h and 24h,sampled and observed the cell morphology under the microscope;collected the cells and extract the protein.Detect the expression of all5278 protein during different times by Western Blot;collected the cells and extract the R-NA.Detect the expression of all5278 protein during different times by RT-PCR.1.2.3 ResultsBioinformatics analysis:the full length of all5278 gene is 1131bp,encoding a pr-otein with 376 amino acid residues,and its molecular weight is 42302.0Da,all5278 b-elongs to PKc-like superfamily,it’s a hydrophilic protein.The tertiary structure of al-15278 is similar with pknG and CDC2.Using the CDC2 inhibitor roscovitine examined the relationship between cyan-obacterial cell viability and expression of all5278:roscovitine promoted the growth of Synechococcus sp.PCC 7002 after treated 4 hours,and had a significant dose-dep-endent inhibition on the hydrogenase activity of Anabaena sp.PCC 7120.The effects on Synechocystis sp.PCC 6803 was not observed.After roscovitine treated 4 days,t-he growth and viability of PCC 7120 were significantly decreased;the growth of PC C 6803 was also shown significant inhibition compared with the control group althoug-h its vitality showed little difference.The growth and viability of PCC7002 were not affected after the 4 days treatment.Microscopic observation showed that roscovitine d-id not affect cell morphology of the three cyanobacterial strains,but shorter filaments of PCC7120 was observed after 4 days treatment.Induce Anabaena sp.PCC 7120 to enter cell division by using red light,exa-mined the correlation between the expression of all5278 with cell division:After c-ultured in the nitrogen-shortage BG-11 medium,and induced by red light,PCC7120 e-nter the cell division cycle.Analysis found that the expression of all5278 protein was significantly higher under red light stress after treated 5 and 24 hours.The results sho-w that the transcription of all5278 gene was significantly higher under red light stress after treated 2,8 and 24 hours.