淡色库蚊氯菊酯抗性相关基因PR-XP1 cDNA的克隆与序列分析

【目的】克隆淡色库蚊一个全新的氯菊酯抗性相关基因PR-XP1的全长序列,分析、预测其编码蛋白的结构与功能,为阐明该基因的抗性机制及研制新型卫生杀虫剂奠定基础。【方法】依据抗性与敏感品系库蚊差异表达的EST片段（GenBank号:EC093826.1）,设计特异性PCR引物,采用RACE技术,克隆淡色库蚊氯菊酯抗性相关基因PR-XP1的全长cDNA序列,运用生物信息学软件对其同源性与系统进化进行分析,并对PR-XP1编码蛋白的结构与功能进行预测。【结果】获得了全长为2 004bp的淡色库蚊氯菊酯抗性相关基因PR-XP1,该基因具有完整的CDS区,编码249个氨基酸。同源性比对与系统进化分析表明,PR-XP1基因核苷酸序列与致倦库蚊（GenBank号:XM₀01862469.1）、埃及伊蚊（GenBank号:XM₀01658032.1）和冈比亚按蚊（GenBank号:BX021208.1）相关基因的同源性分别为95%,83%和70%;其编码蛋白的氨基酸序列与致倦库蚊、埃及伊蚊、西方蜜蜂、入侵红火蚁、佛罗里达弓背蚁、印度跳蚁等昆虫中"假设性蛋白"的氨基酸序列同源性均达46%以上。结构分析显示,PR-XP1基因编码蛋白可能为具有2个跨膜螺旋的膜蛋白,跨膜区形成了一个特异性的"U"形结构;该基因还具有1个微弱的信号肽位点和24个磷酸化位点。【结论】克隆得到了一个全新的淡色库蚊氯菊酯抗性相关的"保守性假设蛋白"基因PR-XP1,推测其功能与氯菊酯抗性相关。     

Effect of dietary GroBiotic®‐A supplementation as a prebiotic on the intestinal microflora,growth performance,haemato‐serological parameters,survival rate and body composition in juvenile beluga (Huso huso Linnaeus, 1754)

The objectives of the present study were to investigate of GroBiotic^®‐A (GBA) on growth, autochthonous intestinal microbiota and haemato‐serological parameters of beluga juvenile. A total of 180 fish (40.82 ± 5.81 g) were fed diets containing graded levels of GBA (0, 5, 10 and 20 g kg^?1 diet) for 8 weeks. No significantly differences in body composition, total viable aerobic bacteria, mean cell volume, mean cell haemoglobin, mean cell haemoglobin concentration, neutrophils, eosinophils, monocytes, albumin, glucose, triglyceride, cholesterol, alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase and lactate dehydrogenase were detected between treatment groups. The fish fed on 10 and 20 g kg^?1 GBA significantly showed higher lactic acid bacteria, final weight, body weight increases, weight gain and lower feed conversion ratio compared with the control and 5 g kg^?1 groups. The group fed 20 g kg^?1 GBA showed a highly significant difference in condition factor, survival rate, final length, total red and white blood cells, lymphocytes, haematocrit, haemoglobin, total protein, total immunoglobulin and lysozyme activity. The specific growth rate of the treatment groups was significantly elevated compared with the control groups. These results indicated that GBA at level 20 g kg^?1 improved growth, welfare and survival of beluga juvenile.     

Feed consumption, nutrient utilization and serum metabolite profile of captive blackbucks (Antelope cervicapra) fed diets varying in crude protein content

A feeding trial was conducted to determine the optimum level of crude protein (CP) in the diet of captive blackbuck (Antelope cervicapra) in which feed consumption and nutrient utilization are maximal. Fifteen blackbucks (BW 25-34 kg) were distributed into three groups of five each in an experiment of 75-days duration including a digestion trial of 5-day collection period. All the animals were offered 200 g of concentrates and fresh maize fodder ad libitum. The overall CP content of the three respective diets was 6.9%, 10.4% and 12.7%. Blood samples were collected on the last day of the experiment. Intake and digestibility of CP increased (p < 0.01) with the increased level of CP in the diet. Feed consumption and nutrient intake were not significantly different among the groups. However, digestibilities of most of the nutrients were higher in the 10.4% CP diet than in the 6.9% CP diet. The endogenous loss of nitrogen was similar among the groups. Based on the endogenous losses, minimum N requirement was calculated to be 776 mg/kg BW(0.75) /day, and to meet this requirement, diet must contain at least 8.27% CP. Serum urea nitrogen concentration increased (p < 0.01) with increased level of dietary CP. Serum level of glutamate oxaloacetate transaminase and glutamate pyruvate transaminase was higher (p < 0.05) in the group fed 6.9% CP diet. Animals in the group fed low protein diet also lost body mass during the experimental period. It was concluded that a diet containing 10.4% CP was optimum for maximizing nutrient utilization without any adverse effect on voluntary feed consumption and serum metabolite profile of blackbucks.     

绿盲蝽化学感受蛋白基因AlucCSP1的克隆、表达谱分析及结合特征

 【目的】克隆、组织特异性表达以及荧光结合特征分析绿盲蝽 (Apolygus lucorum Meyer-Dür)化学感受蛋白（chemosensory protein,CSP）。【方法】设计特异性引物,用RT-PCR技术克隆绿盲蝽化学感受蛋白基因AlucCSP1,使用半定量RT-PCR和Real-time PCR技术对AlucCSP1组织特异性表达进行研究,在BL2（DE3）工程菌株中用pGEX-4T-1载体进行原核表达,并在GSTrap FF柱中纯化和去标签蛋白。使用bis-ANS作为荧光配基,研究AlucCSP1蛋白与58种气味标样和5种棉花次生代谢物质的结合特征。【结果】得到了绿盲蝽化学感受蛋白基因,命名为AlucCSP1（GenBank登录号：JN573217）。该序列阅读框全长393 bp,编码130个氨基酸残基,N端有一段长17个aa的信号肽,具有4个保守的半胱氨酸残基。绿盲蝽AlucCSP1几乎全部在雌虫触角中表达。构建pGEX/AlucCSP1原核表达载体,在BL21（DE3）工程菌株中表达出分子量约为39 kD的融合蛋白,用亲和层析法纯化、经凝血酶作用去除标签蛋白获得纯化的AlucCSP1蛋白,该蛋白与棉酚、单宁、槲皮素、芦丁水合物亲和力较强,结合常数分别为13.4、32.7、19.7、38.5 μmol•L-1。【结论】克隆得到绿盲蝽化学感受蛋白基因AlucCSP1,在雌性触角内特异表达,并进一步得到约13 kD的AlucCSP1纯蛋白。为研究AlucCSP1在绿盲蝽化学感器中的作用奠定了基础。     

血粉膨化机的设计

禽畜血液富含粗蛋白，目前常用的加工方法如喷雾干燥法，发酵法等存在产品消化率低，投资较大等问题，膨化处理能提高蛋白消化率。但血粉比淀粉类原料难膨化，存在着设计上的特殊性。本文在理论分析及试验结果的基础上，对膨化机的特殊结构及尺寸设计进行了分析。实际应用表明该设计基本解决了技术及工艺问题。     

作物害虫的猖獗与高毒农药的"倾泻"的治理对策

阐述了我国农作物害虫发生现状及化学农药防治的现状，分析了作物害虫猖獗与高毒农药“倾泻”的原因与发展，提出了防止作物害虫猖獗和高毒农药“倾泻”的5条对策，即做好高毒农药危害性的宣传工作，提高农民禁用、限用高毒农药的自觉性；调整农药产品结构，大力发展生物农药；生物农药与化学农药复配，促使高毒农药低毒化；推广转基因作物；推广使用高效、高选择性、低毒、低用量、安全农药。     

壳聚糖对大豆蛋白发酵液的絮凝研究

对用壳聚糖絮凝大豆蛋白发酵液进行研究,讨论pH值、絮凝剂浓度、温度和搅拌时间对絮凝效果的影响,结果是壳聚糖用量200 mg/L、pH值为4、搅拌12 min、温度30℃时效果最好.     

中国荷斯坦牛SRY基因编码区的克隆与原核表达

利用PCR技术从雄性中国荷斯坦牛的基因组DNA中克隆了Y染色体性别决定基因(Sex-deter-m in ing R eg ion on the Y Chrom osom e,SRY)的编码区全长序列,构建了表达载体pET-28a/SRY,并将其在大肠杆菌(E.coli)中进行了诱导表达,对表达产物进行了检测。结果表明,SRY基因编码区长687 bp,编码229个氨基酸;表达载体pET-28a/SRY构建成功;表达产物中含有相对分子质量为33 ku的SRY蛋白。     

转抗虫基因杨树对土壤微生物影响分析

以4年生转双抗虫基因741杨试验林为对象,研究了土壤中Bt毒蛋白的分布,及其在生长季节中土壤微生物种群数量的变化.结果表明:试验林土壤中发现了毒蛋白的存在,在转基因植株根围含量最高,距根系越远,含量逐渐降低.其毒蛋白分布趋势是根组织＞根际土壤＞根围土壤＞地表土,其浓度一般呈10倍的等级降低,在非转基因对照植株根系周围未...     

猪粪堆制过程中铅、镉、铜、锌的变化

对猪粪在高温好氧堆制过程中铅、镉、铜、锌全量以及相对含量随时间的变化进行了深入研究.结果表明:猪粪中铅、镉、铜、锌总浓度随着堆腐时间的延长呈不断增加趋势;铅、镉、铜、锌的相对含量呈现先增加后下降的趋势;堆腐温度和pH是影响DTPA-Pb、Cd、Cu和Zn的相对含量变化的主要因素.