A genome project has been launched and aims to sequence total genome of Cotesia plutellae bracovirus. On this process, several open reading frames (ORFs) have been identified. This study was intended to clone and express protein tyrosine phosphatase genes, PTP1 and PTP6. The ORFs of these two genes consist of 900 and 891 bp, respectively. PTP1 and PTP6 are genes of a group of genes that has been implicated as important regulatory components in cell growth, differentiation and malignant transformation by certain viruses. In this work,we studied the cloning and expression patterns of these genes in Plutella xylostella, a lepidopteran host of C. plutellae. A polymerase chain reaction (PCR) produced the corresponding products of PTP1/6. These PCR products were cloned and expressed using an expression vector pBAD-TOPO, and then over-expressed using an inducer, L-arabinose. Then the purified proteins were confirmed using Western blotting (immunoblotting using V5 antibody) and the apparent molecular weights of both proteins were about 40 kDa. Expression of PTP genes were analyzed in the parasitized P. xylostella by realtime RT-PCR, which indicated late expression pattern of both PTPs during parasitization. We are pursuing to elucidate their physiological function in the parasitized host insect.

This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.