Cloning and Expression of HBV Infection Related Novel Gene C12orf49

Objective To clone, express and purify C12orf49 recombinant protein. To prepare rabbit anti-C12orf49 protein polyclonal antibody in order to further elucidate its biological function. Methods PCR was used to amplify the gene C12orf49 in vitro. pET-32a (+)-C12orf49, the recombinant protein prokaryotic expression vector, was transformed into E. coli. IPTG was used as the inductive agent to obtain C12orf49 recombinant protein, and the recombinant protein was analyzed with sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and Western blot. Specific polyclonal antibody was derived from rabbits that immunized by recombinant protein. ELISA and Western blot were used to test its titer and specificity, respectively. MTT cell proliferation experiment was carried out to observe effect of the protein on proliferation of HepG2 cells. Results The C12orf49 recombinant protein was expressed in a large quantity. Data of ELISA indicated that the titer of polyclonal antibody was higher than 1:1 280 000. And the antibody also had a good specificity, confirmed by Western blot. C12orf49 recombinant protein may had a advanced effect on the proliferation of HepG2 cells. Conclusions Using C12orf49 recombinant protein, we can obtain the polyclonal antibody with great titer and good specificity. Human novel gene C12orf49 encoded protein could promote the proliferation of HepG2 cells.

Parasite-induced learned taste aversion involving Nippostrongylus in rats

SUMMARYAfter demonstrating that rats were capable of discriminating between the same diet treated with either flavour 1 or flavour 2 and that the 2 flavours of diet were equipreferred, an experiment was carried out to see whether learned taste aversion might play a role in the reduction of food intake that is commonly observed during the course of a parasitic infection. The results showed that rats, given a subcutaneous inoculation of approximately 6000 third-stage larvae of Nippostrongylus brasiliensis (Nematoda) while feeding on a diet containing flavour 2, strongly preferred to eat diet containing flavour 1 when given simultaneous choice conditions. Uninfected rats showed no preference and ate equal amounts of both flavoured diets. This effect is interpreted as the first experimental demonstration of learned taste aversion using a eukaryotic parasite as the inductive agent.

The induction of cytoplasmic petite mutants of Saccharomyces cerevisiae by hydrostatic pressure

This study demonstrates that hydrostatic pressure is a potent inductive agent of the petite mutation in cultures of Saccharomyces cerevisiae. The inductive capacity of this mutagen is dependent on the magnitude and the duration of the pressure treatment. Furthermore, the extent of petite induction varies with the growth stage of the culture. Induction occurs in pressure-treated (1-4 X 1-(4) lbf in.-2 or 9–66 X 10(4) kN m-2 for 4 h) log growth cultures but not in stationary or lag phase cultures. Petite induction and cell survival are also dependent on the particular strain of yeast which is pressure-treated. Tetrad analysis and complementation assays demonstrate that pressure-induced petite cells are cytoplasmic in nature. Moreover, induced petite cells show a wide range of suppressivity (2–99%) with a large proportion of the petite cells being highly suppressive.