Effect Of Somatostatin Genetic Engineering Subunit Vaccine On Growth Of Mice

In order to construct p ET-32a-2SS28-SS14, and p ET-32a-3SS28 recombinant prokaryotic expression plasmids, two kinds of Somatostatin gene(SS14 and SS28 gene) were cloned into p ET-32 a vector. The expressed protein was used as subunit vaccine to vaccinate mice, to study their effects on the growth of mice, and investigate the different effect of Somatostatin subunit vaccine on different gender mice, and to study the effect on growth of different animals, in order to improve the economic efficiency of animal production.1 Construction of prokaryotic expression vector of p ET-32a-2SS28-SS14 and p ET-32a-3SS28In order to construct p ET-32a-2SS28-SS14, and p ET-32a-3SS28 recombinant prokaryotic expression plasmids, two kinds of Somatostatingene(SS14 and SS28 gene) were cloned into p ET-32 a vector. PCR amplification and sequencing showed the target gene inserted in the p ET-32 a plasmid,the sequence and the direction were correct.2 Expression and purification of 2SS28-SS14 and 3SS28 proteinp ET-32a-2SS28-SS14 and p ET-32a-3SS28 recombinant plasmids were transform into E. coli BL21 De3, and the target protein expression was induced by IPTG, and western blot analysis showed the correct expression of target protein. After cell sonication,centrifugation and protein solubility analysis, the supernatant was purified.3 Preparation of myostatin subunit vaccine and animal immunationThe subunit vaccine was made of purified target protein and adjuvant emulsion. 60 Kunming mice(4 weeks old, half male and half female) were selected and immunized. Male and female mice were randomly divided into 3 groups, 10 mice in each group, every mouse was marked. The group as follows: 10 female and 10 male mice were immunized with 2SS28-SS14 protein; 10 female and 10 male mice were immunized with 3SS28 protein; 10 male and 10 female mice were immunized with white oil emulsion solution as negative control. The vaccine immune group of each mouse was immunized with 100 ug protein. The control groups were injected with equal dose of oil emulsion. After 1 weeks of immunization, each group was second immunized. Each mouse immunization with 50 ug protein, control group was immunized withequal dose of oil emulsion. During 4 weeks to 7 weeks, every mouse was weight once a week. SPSS software was used to perform the statistical analysis. Compared with the blank control group the average weight gain of female mice in experimental group of two kinds of vaccine, the difference was not significant(P>0.05). The average weight gain of male mice immunizatedwith 3SS28 was not significant(p>0.05)in the first and second weeks, compared to blank vaccine control group;however, the average weight was significantly higher than that of control group(P<0.05) after third week. The average weight gain of male mice immunizated with 2SS28-SS14 was significantly higher than that of control group(p<0.05), in the first and second weeks after immunization, the weight difference was very significant(P<0.01) in third week. That differentsubtype myostatin subunit vaccine has different effects on the growth of different gender Kunming mice; from the increase of body weight of male mice, molecular subtype SS-14 was better than SS-28 subtype, indicating the immunogenicity of myostatin of different subtypes is different.In summary, recombinant p ET-32a-2SS28-SS14 and p ET-32a-3SS28 prokaryotic expression plasmid were constructed successfully by gene cloning technique in this study and also detected their expression in E. coli. And the expression of the target protein was purified and vaccinated mice, the average weight gain(P<0.05)of male mice immunizated with3SS28 b was significantly higher than that of blank group, the average weight gain of male mice immunizated with 2SS28-SS14 was higher than that of male mice immunizated with 3SS28(P<0.01). In female mice test the two vaccines havn’t obvious difference. Therefore, the efficient Somatostatin subunit vaccine was obtained in this study and laid the foundation of basic research and application of Somatostatin subunit vaccine.