Cloning And Sequence Analysis Of Ankyrin And PTP Genes And Identification Of A Ankyrin Promoter

Cotesia vestalis Bracovirus (CvBV) plays a key role in the interaction between Cotesia vestalis and its host, Plutella xylostella L. Ankyrin and PTPs (protein tyrosine phosphatases) encoded by CvBV are the largest families that the CvBV encoded. The large number of members and their early expression reflect their important roles in parasitism, especially in suppressing insect immune cells. The study of the structure of the genes and their promoters may be useful for the understanding of the mechanisms of the immune suppression on the hosts, especially in signaling pathway, such as NFkappaB signaling pathway. The results are summarized as follows:1) The full cDNA sequences of seven genes belonging to two gene families Ankyrin and PTP were obtained using Rapid Amplification of cDNA Ends (RACE). They were designated as CvBV-C2-2, CvBV-C12-2, CvBV-C12-5, CvBV-C12-11, CvBV-C14-5, CvBV-C14-6and CvBV-C28-1, which were966bp,1130bp,1301bp,1245bp,1383bp,1177bp and1057bp long respectively. The full cDNA sequences consisted of open reading frames of492bp,900bp,894bp,948bp,972bp,1005bp and831bp long encoding163,299,297,315,323,334and276amino acid residues respectively. The seven predicted mature enzymes had molecular masses of18867.54,34607.06,34514.32,36583.69,36583.69,39166.89and32001.44Daltons.2) The promoter structure of the gene CvBV-C17-4was analyzed. The upstream sequences,-lbp～-1013bp,-1bp～-1750bp,-1bp～-2902bp(named as C17-4P1, C17-4P2, C17-4P3) were truncated, and then respectively inserted into pGL3-Enhancer Vector, which contained firefly luciferase reporter. After transfused by the above vectors, HEK293cells were cultured. The relative light intensity of fluorescence for the samples was detected. The values of sample C17-4P2, and C17-4P3were easily measured, though the former is significantly higher than the latter. However, the value of the sample C17-4P3could be ignored.