Cloning And Expression Of The Vp39 Gene Of SeMNPV And The Research Of Surface Plasmon Resonance Biosensor As Well As Applying

1. Cloning and expression of the vp39 gene of SeMNPV Because of its huge potential value in the fields of the construction of high efficiency expression vector and the development of reconbinant virus pestcide ,insect baculovirus(IBV) becomes one of the hotspots of molecular biology research. Nuclear polyhedrosis virus(NPV) is the most important genus of IBV with nearly 447 species. Nuclear capsid protein gene(vp39) was proved to be closely related to the infection process.A fragment of SeMNPV (Spodoptera exigua multiplenucleocapsid nuclear polyhedrosis virus) about 1 .3kb had been amplified by employing PCR technique and by designing a pair of primers based on the sequence of capsid protein gene vp39 of AcM7NPV(Autographa caizfornica multiplenucleocapsid nuclear polyhedrosis virus). The recombinant expression plasmid pET-Se39 has been constructed by subcloning the 1.3kb fragment into the procaryotic expression vector pET-28. After transformed into E. coil BL2 1, SeMNPV vp39 gene was highly expressed by the inducement of IPTG. SDS-PAGE analysis showed that the molecular weight of the expressed protein was about 39kD. The amount of the expressed product reached to the highest level when it had been induced with IPTG for 4 hours. The encaryotic expression plasmid pGEM-IE1-Se39 containing an iel promotor has been constructed. This research would redound to the deep study of the structure and the function of the vp39 gene ,and furthermore, would be helpful to the research of the relation between virus and hosts. 2. Research and applying of surface plasmon resonance biosensor Biomacromolecule interaction is so important a mechanism that nearly every proceeding in the body of creature, the copy of gene,the transcription of gene, the expression of gene, the enzymatic reaction,the transmission of signal, etc., related to interaction of bioniacromolecule. Surface plasmon resonance(SPR) biosensor is a novel biomacromolecule interaction detection instrument. Compared with other methods, surface plasmon resonance biosensor has many unique advantages. Not only is it convenient and sensitive, but also it is unnessisary to mark samples with enzyme, fluorescence marker or radioactive tracer. With several sorts of biological products e.g. hepatitis B surface antigen/antibody and tetanus toxsoid/antitoxm,, the speciality of surface plasmon resonance biosensor has been discussed in the paper.