The Studies On The XNA Microarray, HaSNPV Bacmid And HzAM1 RpL21 Gene

There are four chapters in this thesis. Chapter 1 is a general introduction of biochips,electrochemical analysis, baculovirus,Cre/LoxP system and ribosome. In chapter 2 a novel electrode was developed based on the XNA on Gold? microarray platform. The platform combines self-assembling monolayers. Electrochemical analysis revealed the ferrocene redox current for the SAMS modified electrode was greater than that with bare gold electrode. The electrochemical reaction of K4Fe(CN)6 was reactivated upon addition of ferrocene on SAMS coating. Addition of DNA to the SAMS resulted in only a minor change in the electrochemical signal. Experiments proved that the coating has excellent stability. These studies show that the microarray platform and electrode can be used to analyze different samples. Chapter 3 was about the construction of a LoxP-containing HaSNPV bacmid. The 3'RACE results showed that the region between the 3' ends of ORF6 and ORF7 was a right place for bacmid cassette insertion. The SV40-lacZ gene of pBluelox was changed by hsp70-lacZ gene, which can function in insect cells, generating plasmid pBluelox1. The BamHI site in pBluelox1 was removed and a plasmid pBluelox2 was generated. The ORF6 and a LoxP containing ORF7 were cloned into pBluelox2, resulting in the transfer vector pBlueLZP. Now the co-transfection of pBlueLZP and HaSNPV to HzAM1 cells is in process.In chapter 4 RpL21 gene of HzAM1 cell was cloned and its sequence was analyzed. The length of this gene is 775bp, which include a 295bp intron. This gene encodes a 159 amino acid protein, whose predicted molecular weight and isoelectric point are 17.97 kDa and 10.996 respectively. There are 3 possible phosphorylation sites and 1 glycosylation site in the RpL21. Homologue comparison result indicated that RpL21 was highly conserved. There were two highly conserved regions in the RpL21 alignment, and the N-terminal part is more conservative than the C-end. The phylogenetic tree based on RpL21 sequence was representative to the phylogeny relationship between the species.