哺乳动物嵌合体制作及分析进展

嵌合体技术对研究哺乳动物早期胚胎的发育,探索细胞分化规律,掌握基因的表达调控规律,建立遗传疾病动物模型等具有重要意义。本文分别就近几年来嵌合体动物的制作方法及分析手段作一简要概述。     

苜蓿秋眠性研究进展

苜蓿Medicago sativa的秋眠性实质上是在秋季日照变短和温度降低时植株所表现出的一种适应性生长特性,这种生长特性与植物的抗寒性和生产性能相关.早在20世纪20年代,美国就发现了这一现象,但直至20世纪70年代末期,随着秋眠性测定方法的发明和分类标准的制订,才使它在苜蓿生产中发挥着越来越重要的作用,在许多国家秋眠性已成为栽培选择品种时的首要指标.概述了秋眠性的概念及发展史、测定方法、有关生理生化研究进展及其在国内外的应用.     

Isolation and Identification of a Variant Porcine Epidemic Diarrhea Virus and the Immune Efficacy of Its Inactivated Vaccine

This study was aimed to isolate a mutant strain of porcine epidemic diarrhea virus and prepare a PEDV inactivated vaccine with high valence by suspension culture process for immunizing against PEDV effectively in China.200 small intestines and theirs contents of diarrhea piglets died of diarrhea,collected from many large-scale pig farms in China,were detected by RT-PCR and sequenced,a mutant strain of porcine epidemic diarrhea virus was selected and put on the suspension-cultured Vero cells in a 2 L reactor for virus isolation and continuous cell culture,the harvested virus suspension,which was identified and determined TCID₅₀,was inactivated by formaldehyde and mixed with aluminum hydroxide gel adjuvant to prepare PEDV inactivated vaccine.After its physical behavior,stability viscosity,sterility test were checked out,the safety and immune efficacy were studied by immunizing the pregnant pigs and theirs piglets.The results were as follows:86 samples were detected positive in 200 samples,cytopathy occurred after the mutant strain samples screened were passaged to 5th generation,the virus suspension was harvested in 10th generation and identified as a mutant strain of PEDV,named PEDV-GF10 strain.The virus titer of harvested virus suspension was measured up to 1×10^8.0 TCID₅₀/mL after concentrated.After the vaccine was checked out,the sows,40 and 25 days before delivery in experimental groups,were injected into Xuehai acupoint with 4 mL vaccine and the pigs in blank group were free of immunifications.The results showed that there were no obvious differences in the production status of the sows in experimental groups and blank group and the temperature of theirs 3-day-old healthy piglets injected different doses of vaccine,and the vaccine was safe to the sows and piglets.Forty 3-day-old piglets producted by pregnant sows in experimental groups and blank group were randomly selected and taken 4 mL 1×10^8.0TCID₅₀/mL F10 virus culture.The PEDV morbidity of piglets in blank group was 100% after injection and the antibodies were negative;10% piglets in blank group had mild diarrhea symptoms,the protection rate was up to 90%,antibody of passive immunity in piglets lasted for more than 35 days.Virus titer of mutant strain of PEDV-GF10 improved a lot by suspension cell culture,the PEDV-GF10 inactivated vaccine was safe,and could effectively prevent and control the variation strain of PEDV in China.     

甘肃羌活资源特征及开发利用

在多点、多次实地考察和查阅大量文献资料的基础上,对羌活Notopterygium incisum生物学特性、品种分类以及在甘肃的资源分布状况作了较系统的介绍,并对其开发利用进行了探讨.     

基因枪轰击不同剂量小鹅瘟病毒VP3基因疫苗在雏鹅体内的动态分布

本文开展了检测小鹅瘟病毒(GPV)VP3基因的实时荧光定量PCR(FQ-PCR)方法的建立和基因枪轰击不同剂量(1、3和6μg)GPV-VP3基因疫苗(pcDNA-GPV-VP3)在30日龄四川白鹅体内(心、肝、脾、肺、肾、法氏囊、胸腺、哈氏腺、十二指肠、空肠、回肠、直肠、盲肠、胰腺、血液、脑及注射部位皮肤)分布规律的研究。结果表明:①建立的FQ-PCR特异性强、灵敏度高、重复性好,核酸模板数与FQ-PCR测定的Ct值相关系数达到0.999,具有很好的直线相关性;②pcDNA-GPV-VP3各剂量免疫雏鹅1 h即可在各组织中检测到,其中注射部位含量最高,肝、肾、淋巴器官(脾、法氏囊、胸腺、哈氏腺)含量较高;③到免疫后217 d时,1μg组免疫雏鹅各个组织器官内仍检测到pcDNA-GPV-VP3的存在,但多数组织器官中的含量比1 h时约少了4个数量级,其中免疫部位减少了7个数量级;④血液中pcDNA-GPV-VP3的含量较少,且免疫后1 h~217 d各时间点的差异不显著(P≥0.05);⑤不同剂量pcDNA-GPV-VP3免疫雏鹅各组织中的含量呈现的总体规律为6μg组>3μg组>1μg组,但差异不显著(P≥0.05)。因此,FQ-PCR是定量检测pcDNA-GPV-VP3在免疫雏鹅体内含量的可靠方法,pcDNA-GPV-VP3免疫雏鹅后1 h时可分布至雏鹅体内各组织器官中并持续存在217 d以上。     

用LSAB免疫组化染色法检测猪生殖-呼吸道综合征病毒抗原的研究

本研究在国内首次成功建立了辣根过氧化物酶标记的链霉亲和素-生物素（LSAB）免疫组化染色法检测猪生殖-呼吸道综合征病毒（PRRSV）抗原。应用LSAB染色技术检测12头人工感染PRRSV美洲株（ATCCVR-2332）或国内分离株（B96-4,B96-5）的SPF仔猪组织细胞内的PRRSV抗原,阳性检出率为100％。LSAB免疫组化染色法操作简单、灵敏度高、特异性强,结果清晰、稳定、重复性好,是一种检测组织细胞内PRRSV抗原的有效方法。     

盐胁迫对牧草种子萌发及其恢复的影响

观察了７种牧草种子在８个ＮａＣｌ浓度;０（０Ｍｐａ）,２１－９－０．１Ｍｐａ）,６６（－０．３Ｍｐａ）,１１ｐ（－０．５Ｍｐａ）,１５０（－０．７０Ｍｐａ）,２００（－０．９Ｍｐａ）,２６７（－１．２Ｍｐａ）,３３４（－１．９５Ｍｐａ）,４４５（－２．６０３Ｍｐａ）ｍｍｏｌ／Ｌ下的发芽率及胚根和胚芽的生长,将盐胁迫下未萌发种子移入营养液中,观测它们在解除发芽率呈下降趋势,但长穗偃麦草的发芽率几乎不     

在语文教学中如何培养学生的“创新精神”？

创新是一个民族进步的灵魂,是国家兴旺发达的不竭动力源泉。因此,对创新人才的培养,应该从基础教育抓起。为了在语文教学中培养学生的“创新精神”,教师首先应转变观念,使自己成为有创新精神的人。其次为学生创设和谐、民主的学习氛围,让学生最大限度地发展创造性思维能力。应让学生丰富知识、夯实基础、拓展视野,为创新思维的发展打下坚实的基础。     

16S rRNA甲基化酶rmtB在猪肠道菌中的传播方式分析

为研究16S rRNA甲基化酶rmtB在猪肠道菌中的传播方式,用脉冲场凝胶电泳(PFGE)分别对来源于A、B猪场的48株rmtB阳性大肠埃希氏菌进行基因分型;用膜接合试验研究rmtB基因的水平传播方式;用微量稀释法对rmtB阳性菌及其接合子进行药敏试验。有45株rmtB阳性大肠埃希氏菌(45/48)能进行PFGE分型,可分为28种不同的PFGE基因型。其中来源于A猪场的20株菌可分为12种基因型,来源于B猪场的25株菌可分为16种基因型;46株rmtB阳性菌可以通过接合试验将rmtB基因及其介导的耐药性传递给受体菌E.coliC600和E.coli488 Rifr,接合频率从4.6×10-13~3.0×10-6不等,接合子对卡那霉素、阿米卡星、妥布霉素、西梭米星、萘替米星、庆大霉素6种二脱氧链霉胺类抗生素均高度耐药(MIC≥512μg/mL)。结果表明,rmtB基因位于接合性质粒上,该基因在两猪场的大肠埃希氏菌中既存在克隆传播,又存在水平传播,以水平传播为主。     

猪表皮生长因子基因的遗传变异及其与产仔性状的相关性分析

本研究通过建立两次PCR扩增分型法,分析猪表皮生长因子(epidermal growth factor,EGF)基因第3内含子上的875bp插入片段在13个中外不同繁殖性能的猪种中的遗传变异,以及EGF基因在白色杜洛克×二花脸资源家系180头F2母猪中与繁殖性状的相关性。结果表明,不同类型中国地方猪种与西方商业猪种在EGF基因位点上存在偏态分布。在资源家系中该位点对总产仔数、产活仔数和断奶活仔数无显著影响(P0.05);尽管BB基因型的产活仔数和断奶活仔数分别比AA型多1.74和0.46头,但BB型的个体太少,各基因型间的均值差异不显著(P0.05);虽然产活仔数的a值和d值分别达到0.87和1.24头,但该基因位点的加性效应和显性效应并不明显。因此,需要利用更多的SNP位点和更多样本进一步了解EGF基因与繁殖性状的关联性。