H90,ferritin,CuZnSOD Gene Clone With Expression Analysis Of Galaxea

In recent years,the coral ecosystem imbalance problem cause by human activity and natural environment changed have been worse than before,the range of coral reef bleaching are extensive throughout the world.In order to explore the coral resilience mechanism of anti-stress, this article clone and analysis the expression of heat shock proteins(HSP 90)from Galaxea astreata,clone and builtd prokaryotic expression of Ferritin、CuZnSOD Gene from Galaxea fascicularis were obtainde in this study. The main results list next:1.The 2568 bp partial sequence of HSP90 from Galaxea astreata is got through RT-PCR and RACE technology,which including an ATPase functional region, a HSP90 functional region and a polypeptide binding region.To analysis the transcriptional level, three temperature 29℃,32℃ and 35℃ were testd, the results indicate an regular pattern of increased first and drop down then.The reaction time,the maximum expression leverl, and the rate of decrease were affectd by the temperature,the higher temperature being, the longer reaction time required,the bigger maximum expression and the rate of decrease being.2.Utilize normal PCR,this study got a full sequence of Ferritin with 680 bp from Galaxea fascicularis,included an 519 bp ORF zone,encoding 172 amino acids.Ferritin gene prokaryotic expression system were constructed with PET-32 a vector and BL21(DE3) host strain, it can be induced expression a specific 41 kDa fusion protein.Optimization of expression conditions found that the system can be most abundantly expressed fusion protein with 0.7mmol/L IPTG under 32℃,the quantity of fusion protein canl be increased with time,it could available a relatively large amount of fusion protein at 6 hour.3.By the mean of normal PCR,this study got a full sequence of CuZnSOD with644 bp from Galaxea fascicularis,included an 471 bp ORF zone,encoding 156 amino acids. CuZnSOD gene prokaryotic expression system were constructed with PET-32 a vector and BL21(DE3) host strain, it can be induced expression a specific 37 kDa fusion protein.Optimization of expression conditions found the system can be most4.abundantly expressed fusion protein with 0.4mmol/L IPTG under 32℃,the fusion protein will be increased with time,it could available a relatively large amount of fusion protein at 6 hour.