鸡Leptin基因正确性的免疫验证  

Immuno-verification of Correctness of Cloned Chicken Leptin Gene

作　　者：邵西兵[1] 施振旦[1] 于迎春[1] 刘颖[1] 梁少东[1] 陈楠[1] 

机构地区：[1]华南农业大学动物科学学院,广东广州510642

出　　处：《Zoological Research》2005年第2期168-173,共6页动物学研究（英文）

基　　金：国家自然科学基金资助项目(39700102);广东省自然科学基金团队项目资助(04205804)

摘　　要：①取20只240日龄的粤黄鸡种母鸡平分为处理组和对照组,分别在第3、31、63和84d对之接种鸡Leptin重组融合蛋白(处理组)或BSA(对照组)1mL/羽。每天记录产蛋率,每10天测定采食量,每两周称重,在试验的第1、31、53、78和91d采血。结果表明Leptin处理组的血浆抗Leptin抗体水平和Leptin水平在免疫后显著高于对照组(P<0.01);第3和4次免疫后处理组产蛋率比对照组显著低约50%(P<0.05);在第98d试验结束时,处理组腹脂量为(115.60±15.07)g,显著高于对照组[(77.98±5.70)g,P<0.05];试验期间处理组的采食量略低于对照组,增重则略高于对照组,但两者差异均不显著(P>0.05);处理组血浆总T3和总T4水平与对照组差异不显著(P>0.05)。②取12只50日龄的粤黄鸡后备小母鸡,平分成处理组和对照组,分别肌注含抗Leptin抗体的卵黄提取液和生理盐水6mL/只,处理组在2、4、6h的累积采食量显著高于对照组(P<0.05),但在8h的累积采食量与对照组差异不显著(P>0.05)。两个试验通过免疫重组鸡Lep tin融合蛋白,使鸡表现出Leptin功能的丧失,证明目前所克隆的鸡Leptin基因在家鸡中的确存在和其正确性。①Twenty 240 d old Yuehuang hens were equally divided into 2 groups and respectively inoculated i.m. with recombinant leptin or BSA immunogen (containing 1 mg protein in 1 mL mineral oil emulsion adjuvant, n=10 in each group) on days 3, 31, 63 and 84, blood sampled on days 1, 31, 53, 78 and 91 in the experiment. The plasma anti-leptin antibody titres following 1st inoculation became significantly (P<0.01) higher than in the control hens, so were plasma leptin levels,which steady increased following repeated immunizations. Plasma total T_3 and total T_4 levels were slightly lower but not significantly different (P>0.05) in treated hens following repeated inoculations from levels in control hens. The laying rates was significaltly depressed (P<0.05) by leptin immunization by 50% after the 3rd inoculation. Feed intake was slightly lower whereas live weight slightly higher in leptin-immunized than in the control hens. The abdominal fat mass was significantly higher in treated than in control hens (115.60±15.07 g vs 77.98±5.70 g, P<0.05) on day 98. ②Twelve 50 d old pullets were i.m. injected with anti-Leptin antibody in yolk extracts (n=6) or physiological saline (n=6). Pullets passively immunized against Leptin exhibited significantly higher (P<0.05) accumulative feed intake at 2, 4 and 6 h post injection than the saline injected controls. However the total feed intake of the 2 groups were identical at 8 h post-injection (P>0.05). The above results mimicked the biological effects of loss of leptin bioactivities observed in mammals and implied correctness of so far cloned chicken leptin gene, therefore contributed to clarify the disputes regarding this gene.

关 键 词：LEPTIN基因 免疫 采食量 脂肪沉积 产蛋率 粤黄鸡 

分 类 号：S852.4[农业科学—基础兽医学]