蓝藻藻胆体核基因apce的基因工程分析-genetic engineering analysis of phycobilisomes nucleus gene apce in cyanobacteria.docxVIP

AbstractPhotosynthesis is the most important chemical reaction on the earth, and it is the ultimate energy source of any living organisms. Phycobilisome serves as the light- harvesting antenna in cyanobacteria and red algae. These supramolecular protein complexes, are primarily composed of phycobiliproteins, a brilliantly colored family of proteins bearing covalently attached, open-chain tetrapyrroles known as phycobilins.As we know, the energy transformation process is PE→PC→APC→Chla in photosynthesis. Gene apcE is encoding LCM ligation polypeptides which could help the energy transformation of allophycocyanin (APC). The threshold 210bp of the apcE is encoding the part combined with the membrane directly. From the process APC→Chla,the energy transformation is to PSⅠand PSⅡ independently. In particularly, ApcE issupposed a key function gene of energy transformation from APC to PSⅡ.In this work, Molecular biology technique is used to construct plasmids pBT-apcE. mutant pBT-apcE(1~70) was constructed by TaKaRa MutanBEST Kit. The genomic library of PCC6803 is successfully constructed.This paper cotransformates pBT-apcE plasmid and pTRG–apcF plasmid, pBT-apcE (1~70) plasmid and pTRG–apcF plasmid , pBT-apcE plasmid and pTRG library plasmid into the reporter strain. The bacterial two-hybrid system proved that there is no protein-protein interactions in vivo between ApcE and ApcF, but the mutant ApcE and ApcF have interactions in vivo. By applying the bacterial two-hybrid system, this paper explored protein that may have interactions with ApcE in the genomic library. To identify the protein encoded by target DNA, the nucleotide sequence of the target DNA can be determined and compared to nucleotide sequence databases to probe related proteins.Keywords: ApcE; ApcE mutant; ApcF; bacterial two-hybrid system; genomic library目录摘要..................................................................................................IAbstract ...........................................