皖西白鹅催乳素基因克隆、表达及主动免疫后就巢行为分析  

Cloning and expression of goose prolactin and the active immunization effect in Wanxi white goose

作　　者：陈兴勇[1,2] 何艳[1,2] 时丹[1,2] 耿照玉[1,2] 

机构地区：[1]安徽农业大学动物科技学院,安徽合肥230036 [2]安徽省畜禽产业共性技术研究院,安徽合肥230036

出　　处：《扬州大学学报（农业与生命科学版）》2014年第2期33-37,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：安徽省高校优秀青年人才基金资助项目(2012SQRL062ZD)

摘　　要：为研究催乳素(PRL)对地方鹅就巢行为的影响,构建含鹅PRL成熟编码区完整序列的原核表达载体,通过转化Escherichia coli BL21(DE3)建立稳定的原核表达系统,IPTG诱导下获得胞外重组PRL(rPRL)。选成年就巢期皖西白鹅100只,随机分为5组,A组(对照组)注射等量生理盐水,B组口服溴隐亭,C^E组分别颈部皮下多点注射1、2、4mg·mL-1 rPRL。间隔10d进行第2次加强免疫,剂量与首次免疫相同。结果表明:rPRL表达量约262.5μg·mL-1,重组蛋白约占总蛋白质的35.2%。经His-tag柱纯化回收,回收率为64.9%。与对照组相比,首次免疫后C^E组就巢率极显著下降;二次免疫后D、E组就巢率显著低于其他各组;二次免疫10d后各组间就巢率差异不显著,且均上升至20%以上。产蛋率比较表明,首次免疫后各组产蛋率无显著差异;二次免疫后D组产蛋率显著高于C组,但与其他各组差异不显著。rPRL主动免疫可显著降低就巢率、增加产蛋量,但维持时间较短,且鹅群受环境条件影响显著。This experiment was designed to produce a recombinant PRL （rPRL） for use in immunizing Wanxi white geese against PRL and to determine whether these immunizations could decrease broodiness and increase egg production. The complete PRL coding region from Wanxi-white geese was cloned and transformed into Escherichia coli BL21 （DE3） to establish a stable prokaryotic expression system with rPRL obtained extraeellularly through the induction of isopropyl-β-D-thiogalactopyranoside （IPTG）. We found that approximately 262. 5μg ·mL^-1 of rPRL was expressed, accounting for 35.2 % of the total cellular protein. After recycling with a His-tag column, the concentration of purified rPRL showed that the recovery rate of protein was approximately 64. 9% （as determined with Lowry method）. We then randomly divided Wanxi-white geese （n= 100） into 5 groups （n=20 per group） ： A group injected with saline as negative control, B group orally administered with Bromoeriptine as positive controls, C, D and E group subcutaneously injected with 1, 2 and 4 mg · mL^-1 rPRL, respectively. The rPRL injections into multiple points in the neck were considered the first active immunization, whereas rPRL injections similarly repeated 10 days later were considered the second active immunization. After the first active immunization, the broodiness significantly decreased in C-- E group when compared to A group （P〈0. 01）. After the second active immunization with rPRL, broodiness in D and E group was significantly lower than that of other groups （P〈0. 05）. Broodiness showed no difference among groups after 10 d of the second active immunization, however, broodiness was significantly increased to more than 20% in all groups. Egg production percentage showed no significant difference in each group after the first active immunization. While after the second active immunization, egg production percentage in D group showed significantly higher than that of geese in C group （P〈0. 05）, but only numerically highe

关 键 词：皖西白鹅 催乳素 原核表达 主动免疫 就巢 产蛋率 

分 类 号：S835.2[农业科学—畜牧学]