牛GM-CSF与金黄色葡萄球菌黏附素FnBPB共表达质粒的构建与原核表达

采用PCR方法对牛粒细胞-巨噬细胞集落刺激因子(GM-CSF)和金黄色葡萄球菌FnBPB的D区进行特异性的扩增,并通过重叠延伸PCR扩增GM-CSF-FnBPB串联基因,构建了克隆质粒pMD19-GM-CSF-FnBPB。利用表达载体pET-32a(+)对该融合基因片段进行原核表达,SDS-PAGE分析表明,在1mmol/L IPTG诱导浓度下,在39ku处出现了与目的蛋白一致的外源蛋白带,Western blot分析表明,该蛋白具有反应原性,进而证明该融合基因成功在原核细胞中表达。     

牛胚胎玻璃化冷冻技术的发展和应用概况

胚胎冷冻保存是动物胚胎工程的重要组成部分，它是胚胎移植技术在生产中推广应用的重要基础。和现在通用的常规冷冻法相比，玻璃化冷冻具有用时短，所需设备少和操作简单等优点，成为目前国内外胚胎冷冻保存领域发展新方向。本文就牛胚胎玻璃化冷冻的概念、方法和国内外发展状况做一简要综述。     

初探草坪起源与演化,兼论草坪的概念

草坪为一类特殊的草地。草坪起源的关键在于草坪草的分化发生。草坪草、草坪起源及早期演化 ,都是地史时期内 ,特定的气候 -生物 -土壤协同演化 (Coordinateevolution)的产物。移栽自然草坪 ,开始向人工草坪过渡。人工草地的出现 ,通过“重牧”、“刈割”等导致人工草坪的诞生 ;剪草机的发明标志着现代草坪的降临     

肉鸡对三碱基氯化铜、铜氨基酸螯合物、铜蛋白盐生物利用率研究

采用玉米—豆粕型饲粮研究肉公雏对三碱基氯化铜(TBCC)、铜氨基酸螯合物(CuAA)、铜蛋白盐(CuPro)或硫酸铜(CuSO4)的生物利用率。在基础饲粮(Cu14mg/kg,DM基础 ,实测值)上 ,TBCC、CuAA、CuPro、CuSO4 的铜添加水平依次为150mg/kg、300mg/kg、450mg/kg,试验3周。结果表明 ,铜源和铜水平对试鸡体重和饲料转化率的影响不显著。以CuSO4 为参比标准 ,铜添加水平或日粮铜摄入量为自变量 ,经对数转换后肝铜积累为因变量建立多元线性回归模型 ,按斜率法综合求测铜源生物利用率为 :CuSO4 100 % ;TBCC112.4 % ;CuAA127.7 %(P<0.01) ;CuPro99.3 %。结果表明TBCC或CuPro生物利用率与CuSO4同效 ,但CuAA生物利用率比CuSO4 高。该试验提供了评定肉鸡有机铜源生物利用率的实用方法。     

一年生黑麦草在不同基肥条件下的生物量研究

在不同基肥条件下，测定一年生黑麦草的产草量、根系长度和深度，结果表明：有机肥每667m^2 1200kg＋复合肥每667m^2 100kg作基肥，一年生黑麦草的产草量最高，达到53226．6kg/hm^2，根系最发达。     

Comparison Study on Effects of Three Chinese Herbal Medicine on Intestinal Morphology in Weaning Piglets

The objective of this study was to compare the effect of three Chinese herbal medicine (medicated leaven, hawthorn, fructus hordei germinates) on histomorphology of different parts of small intestine in weaning piglets.40 28-day old ternary pigs (Duroc×Landrace×Yorkshire) with the body weight (8.13±1.32)kg were divided into four groups, control group, medicated leaven group, hawthorn group and fructus hordei germinates group, and feeding with 1% Chinese herbal medicine, there were two repeats in each group and each repeat had 5 piglets, slaughtering and sampling at 7th and 14th day morning, respectively, sections were used for HE staining.The staining tissues were taken pictures under a microscope camera (200×) and measured VH, CD, IWT and V/C in different parts of small intestine with Image-Pro Plus.The result showed that the IWH of ileum in medicated leaven group was significantly or extremely significantly higher than control group (P<0.05;P<0.01).The VH and IWT of duodenum and jejunum at 7th day in hawthorn group were extremely significantly lower than control group (P<0.01);The IWT of ileum and the VH of ileum at 14th day were extremely significantly higher than control group (P<0.01).In fructus hordei germinates group, the VH of duodenum and jejunum at 7th day and IWT of jejunum were extremely significantly lower than control group (P<0.01);The CD of ileum at 7th day, the IWT of duodenum and ileum at 7th day were extremely significantly higher than control group (P<0.01).The effects of different simple Chinese herbal medicine on histomorphology of different parts of small intestine in piglets were different, because of the different digestive functions of small intestine, the most effects of Chinese herbal medicine an duodenum, jejunum and ileum were fructus hordei germinates, medicated leaven and hawthorn, respectively.     

新西兰兔肌内和皮下前体脂肪细胞原代培养与诱导分化的研究

为建立新西兰兔前体脂肪细胞的体外培养模型,比较新西兰兔肌内和皮下前体脂肪细胞分化过程中相关基因的差异表达,实验采集 1 日龄新西兰兔背最长肌和皮下脂肪组织,采用胶原酶消化方法,分别从 2 种组织中分离前体脂肪细胞,进行细胞原代培养,绘制细胞生长曲线,并对肌内和皮下前体脂肪细胞诱导分化,利用 RT-PCR 检测相关基因的表达变化。结果表明:细胞在分离后 2 h 已经贴壁,24 h 时呈现出短梭形的细胞形态,第 2 天细胞变成长梭形,第 3 天进入对数生长期。肌内和皮下前体脂肪细胞在诱导分化后均被油红O 染色,皮下前体脂肪细胞的脂质积累在诱导分化的第 2 天显著高于肌内前体脂肪细胞(P<0.05);荧光定量结果表明,肌内和皮下前体脂肪细胞 CCAAT 增强子结合蛋白(C/EBPα)基因的表达趋势在诱导分化过程中相同,肌内前体脂肪细胞过氧化物酶体增殖激活受体(PPARγ)第 4 天的表达量显著高于第 6 天,而皮下前体脂肪细胞 PPARγ表达量差异不显著;肌内前体脂肪细胞脂蛋白脂肪酶(LPL)表达量在第 0天和第 2天差异不显著,而皮下前体脂肪细胞第 2 天显著高于第 0 天(P<0.05);肌内前体脂肪细胞脂肪酸合酶(FAS)基因第 0、2、4 天的表达量差异不显著,而皮下前体脂肪细胞第 2、4 天显著高于第 0 天(P<0.05)。本研究成功构建了新西兰兔肌内和皮下前体脂肪细胞的体外培养和诱导分化模型,并发现皮下前体脂肪细胞分化早于肌内前体脂肪细胞,为进一步研究新西兰兔前体脂肪细胞的分化机制和脂肪沉积奠定基础。     

新牧一号苜蓿MvP5CS基因的克隆和功能分析

为研究苜蓿（Medicago sativa）耐盐分子机制,为抗逆分子育种提供依据,通过同源克隆和RT PCR技术克隆了新牧一号杂花苜蓿（M.varia Xinmu 1）的MvP5CS基因, MvP5CS全长2 148 bp,Genbank登录号为：EU371644。荧光定量PCR分析发现,在盐胁迫下MvP5CS基因表达明显上调,说明 MvP5CS与苜蓿的耐盐性相关。利用农杆菌介导的叶盘转化法,将MvP5CS基因成功转入烟草（Nicotiana tabacum）,盐胁迫下转MvP5CS基因T1代烟草萌发率和根长均高于非转基因烟草。表明新牧一号苜蓿P5CS基因能够提高转基因烟草的耐盐性。     

Acute and Sub-chronic Toxicity Test on Rats of a New Kind of Antidiarrheal Chinese Herbal Medicine Compound Preparation for Livestock

In order to understand the security of a new kind of antidiarrheal Chinese herbal medicine compound preparation for livestock,acute and sub-chronic toxicity test were conducted.Acute toxicity test used the largest drug dose method,20 Wistar rats were orally treated with the Chinese medicine compound preparation.In the sub-chronic toxicity test,80 rats were randomly divided into 4 groups with 20 rats in each group and orally given a dose of 3 000,1 500,750 and 0 mg/(kg·BW）of Chinese medicine compound preparation once a day for 30 days.The general clinical status was observed,rats weight were measured and the dose was adjusted every week during the test,after the test measured blood routine index,biochemistry index,and preceded the gross anatomy observation,weighing each major organs and calculated the viscera coefficient,and proceded main viscera histopathological observation between the high dose group and the control group.The acute toxicity results showed that every rat would be alive gavaged with the lethal dose（LD₅₀）of compound preparation larger 5 g/(kg·BW).The sub-chronic toxicity autopsy showed that except heart,lung,and testicles in individual rats appeared mild bleeding in the high dose group,the other dose group organs found no abnormal change.The haematological index showed except mononuclear cell rate（P<0.05),and hematocrit declined significantly（P<0.05）in the high dose group,all the indexes of the other groups were in the normal range,there was no significant difference from the control group.The test suggested the Chinese medicine compound preparation was no toxicity under the condition of this test according to acute toxicity classification standard of exogenous chemicals by WTO,there was no effect on the growth and development of rats in the sub-chronic toxicity test,and there was no chronic toxicity at least 1 500 mg/kg feeding conditions in short-term repeated application.