家畜繁殖生物技术研究现状及趋势     

张继慈 《西北农业学报》1992,1(2):75-79

牛胚胎移植技术已趋于成熟。我国新疆牧科院用一步细管法移植牛冻胚受胎率达41%;牛和羊鲜胚四分胚移植也产下1头犊牛和6只羔羊。家畜体外受精,因卵子体外成熟和受精卵体外培养尚不过关,目前仍停留在实验室阶段。胚胎性别鉴定,1990年Koopman发现单拷贝基因,该基因是Y染色体的性决定区,命名为SRY,可利用PCR技术制成雄性特异DNA探针盒,进行马、牛、羊、猪早期胚胎性别鉴定。北京农学院等单位用PCR扩增牛SRY序列进行奶牛胚胎性别鉴定准确率达100%。英、日、法等国已获得牛胚胎细胞核移植后代;我国也获得1只核移植兔。北农大和新疆牧科院合作以绵羊精子为载体导入牛生长激素基因rMTbGH DNA成功,外源基因整合率为3%。  相似文献   

Quick detection of sex determining region protein gene in mouse fetuses     

《园艺学报》2003,19(5):622-626

AIM: To detect quickly the Y-chromosome specific sex determining region protein (Sry) gene in mouse fetuses on embryonic day 14.5 with a PCR method. METHODS: We designed specific primers with the OLIGO 5. 0 software. Templates were prepared in 30 minutes by the following way. About 1 mg embryonic tissue but not fetal liver was suspended, and treated with 200μL of lysis buffer, consisting of PCR buffer containing 20 mg/L proteinase K, 0. 5% NP-40, and 0.05% Tween 40, at 60°C for 15 minutes, heated for 5 minutes at 100 °C, 10μL was used as template. The PCR react ion was performed in 50μL, using two sets of primers specific for Sry gene (chromosome Y) and IL-3 gene (chromosome 11) . PCR conditions and cycle numbers were optimized. The assessment of the results was done by electrophoresis in 3% agarose run at high voltage. The specificity of the method was conf irmed by fluorescent in situ hybridization (FISH) using a specific male probe on embryonic tissue cells. RESULTS: Electrophoresis showed that PCR product of male control DNA consisted of a 649 bp product representing the IL-3 gene and a 444 bp product representing the Y-specific Sry gene, female control DNA only one 649 bp product. Fetuses with two bands matching those as seen inmale control DNA are the presumpt ive male fetuses. Fetuses, only the IL-3-associated 649 bp band, are the presumptive female fetuses. These were confirmed by FISH. The ent ire procedure took <3. 5 h. CONCLUSION: The established PCR assay offers a quick, simple, accurate, and sensitive detection of sex determining region protein gene in mouse fetuses. This method allowed the preparation and culture of pure male and female hematopoietic stem cells from fetal tissue.  相似文献   

罹患口腔炎乌苏里蝮的相关生理学指标分析     

姚晨昊  路生鑫  陆宇燕  李丕鹏  薛百强 《野生动物学报》2020,(2):401-406

口腔炎是人工养殖蛇类的常见病之一,为了深入了解和尽早做好预防工作,对罹患口腔炎的成年雄蛇乌苏里蝮(Gloydius ussuriensis)进行解剖观察,并检测了相关器官的系数、血液学指标和激素等。结果表明:整体肥满度、躯体肥满度、肾系数等数值罹患口腔炎蛇略高于健康蛇;而肝系数、精巢系数和脂肪体系数等罹患口腔炎的蛇低于健康雄蛇,且前两者呈显著性差异,后者呈极显著性差异。性激素检测结果显示罹患口腔炎的蛇血液睾酮和雌二醇含量均低于健康蛇,且均具极显著性差异。血细胞指标中,红细胞数量、红细胞分布均差、白细胞总数、淋巴细胞数量和淋巴细胞比率等均为罹患口腔炎的蛇高于健康雄蛇,且具显著性差异,但嗜中性粒细胞和淋巴细胞的比值(NLR)、中性粒细胞数量和比率、中间细胞数量及其比率均为患口腔炎的雄蛇低于健康蛇,且具显著性差异。结论:口腔炎不仅影响了主要代谢器官的器官系数的变化,而且血液多项指标和性激素水平也有明显差异。性激素含量的变化将对乌苏里蝮的正常生殖繁育产生不可忽视的影响。另外,NLR和红细胞分布宽度2个血常规指标可作为早期诊断炎症发生的首选参数。  相似文献   

人工养殖西伯利亚狍肠道微生物组成的雌雄差异     

刘金月  刘德凯  王璐瑶  李响  贾博文  刘艳华 《野生动物学报》2021,(2):428-436

肠道微生物是一个复杂的生态系统,与宿主的健康、疾病以及其他因素有着密切的关系。目前,关于肠道菌群结构和功能的研究较多,但是对西伯利亚狍肠道菌群的研究还未见报道。西伯利亚狍(Capreolus pygargus)属于反刍动物,有独特的消化特性和微生物群来帮助它们适应高纤维的食物。本研究利用高通量测序技术在Illumina MiSeq平台上对14只狍肠道微生物进行16S rRNA基因分析。结果表明:个体样本中门类组成的差异,雌性个体JY3和雄性个体JY9除Firmicutes,Bacteroidetes,Proteobacteria这3类菌群外,其他门类菌群所占比例都很低。在科水平上,雌性个体中Ruminococcaceae在JY7和JY13肠道内含量相对较少,Bacteroidaceae在JY7,JY10和JY13中含量较高。雄性个体中Ruminococcaceae在JY11和JY14肠道内含量较少,Prevotellaceae在JY4中含量较少。在属水平上,雌性个体特有的菌属为Roseburia,Alloprevotella,Lachnospiraceae_incertae_sedis和Coprococcus;雄性个体特有的菌属为Fusobacterium,Succiniclasticum,Streptococcus和Clostridium sensu stricto。Alpha多样性分析表明,Ace指数范围2128.93—772.98,Chao1指数范围2128.89—785.27,Shannon指数范围5.44—3.78,Simpson指数范围0.07—0.01。在不同物种分类水平上,雌雄狍肠道内的优势菌群有些不同。PCoA(principal co-ordinates analysis)分析显示,狍样本几乎混在一起,没有明显差异。  相似文献   

文冠果实时荧光定量PCR内参基因的筛选     

王旭  敖妍  刘阳  张宁  郑雅琪  刘金凤  张行杰  张永明 《分子植物育种》2020,(9):2977-2986

为筛选适用于文冠果(Xanthoceras sorbifolium Bunge)性别分化过程研究和不同器官的内参基因,本研究以文冠果性别分化关键期的顶芽和侧芽、根、茎、叶、种仁、雌能花和雄能花为材料,通过实时荧光定量PCR(qRT-PCR)鉴定7个候选内参基因(Actin,UBQ,EF-1α,TUB,eIF-4α,18S rRNA和GAPDH)在各样品中的表达量,并评价7个基因的表达稳定性。结果表明,7个候选内参基因在不同样品中的表达量存在明显差异,各基因在雌能花中的表达均显著高于其他材料,在雄能花中的表达处于较低水平;适用于文冠果性别分化过程的最优内参基因为UBQ和eIF-4α,适用于不同器官的最优内参基因为EF-1α和GAPDH。研究结果将对今后进行文冠果性别分化过程研究和不同器官中的基因表达分析提供参考。  相似文献   

Thyroid hormones modulate the hypothalamo–hypophyseal–gonadal axis in teleosts: Molecular insights     

I. Swapna  B. Senthilkumaran 《Fish physiology and biochemistry》2007,33(4):335-345

Reproduction in fishes is influenced by thyroid hormones at various levels of gonadal cell differentiation and steroidogenesis. Thyroid hormones have recently also emerged as an important modulator of season- and photoperiod-dependent variations in the reproductive cycle with a possible effect on the hypothalamo–hypophyseal axis and pineal interactions. This review describes the current status of thyroid hormone research in relation to reproduction, with special emphasis on contributions to this field by Indian researchers including our laboratory. Evidence is provided for the multifocal action of thyroid hormones at various levels of the hypothalamo–hypophyseal–gonadal axis affecting reproduction. The underlying physiological and molecular mechanisms pertaining to thyroid hormone modulation of reproduction, such as gonadotropin-releasing hormone (GnRH) synthesis and release, androgen and gonadotropin receptor expression, gonadotropin (GTH) expression, and tissue sensitivity to GTHs are highlighted with relevant discussions of the current technical limitations, applications, and future perspectives of research in this field.  相似文献   

巢式PCR法鉴定猪早期胚胎性别的研究     

彭礼繁  罗光彬  白文林 《畜禽业》2007,(5):14-17

根据猪SRY基因序列设计合成2对巢式PCR引物作为性别鉴定引物,根据猪β-珠蛋白基因序列设计了一对引物作为内标引物建立了猪胚胎性别鉴定的PCR反应体系。公猪可以扩增出187bp的SRY基因片段和255bp的β-珠蛋白基因片段,母猪只能扩增出255bp的β-珠蛋白基因片段。由于巢式PCR只需3～8个细胞就可以在紫外灯下看到扩增结果,而常规PCR则需要较多的细胞,所以胚胎性别鉴定时使用巢式PCR效果更好。  相似文献   

Sex change in coral reef fish   总被引：1，自引：0，他引：1  

Masaru Nakamura  Yasuhisa Kobayashi  Saori Miura  Mohamad Ashraful Alam  Ramji Kumar Bhandari 《Fish physiology and biochemistry》2005,31(2-3):117-122

Gonadal differentiation can take many forms in fish, ranging from gonochorism, where individuals directly develop as male or female and finally possess only testis or ovaries at sexual maturation, to hermaphroditism where the same individuals can produce mature male and female gametes at some time in their lives. Hermaphrodite fish are, thus, an excellent model for studying the plasticity of sex determination and differentiation in vertebrates. We have shown that sex steroids play a principal role in sex differentiation and sex change in fish. Our laboratory implements several fish models that undergo sex change from female to male or male to female or in both directions. In this review, we will briefly discuss recent advances in our understanding of the mechanism of sex change in coral reef fish.  相似文献   

圆斑星鲽sox9基因的克隆与表达          下载免费PDF全文

张乐乐  边力  常青  侯吉伦  陈四清  赵庆  刘琨  葛建龙  刘长琳 《渔业科学进展》2018,39(6):72-80

本研究筛选到圆斑星鲽(Verasper variegatus)的性别相关基因sox9,并通过RACE技术获得了全长序列,基因全长为3287 bp,包括1431 bp的ORF,编码477个氨基酸,368 bp的5¢ UTR和1488 bp的3¢ UTR。在3¢ UTR中有多聚腺苷酸尾和加尾信号AATAAA。通过荧光定量PCR测定了sox9基因在圆斑星鲽成鱼不同组织中的表达水平,发现sox9基因在圆斑星鲽的脑、眼、鳃、心、肝、胆、肠、精巢、卵巢、肾和肌肉等各个组织中都有不同程度的表达。在鳃、脑和精巢组织中检测到较高水平的sox9转录,其中精巢中的转录水平显著高于其他组织,sox9基因在性腺中的表达显示出性别两相性差异。其在精巢中的表达水平要显著高于卵巢,说明sox9基因与雄性性腺发育相关。通过测定sox9基因在圆斑星鲽幼鱼不同发育时期(20、30、40、50、60、70和80日龄)的表达水平,发现其在20~50日龄表达量逐渐下降,在60日龄时表达量上升,推测表达量上升可能与幼鱼性腺分化相关。  相似文献   

利用shRNA真核表达载体干涉雌性鸡胚性腺中芳香化酶基因(CYP19A1)的表达   总被引：1，自引：0，他引：1  

朱晓亮  徐桂云  杨宁  李俊英  郑江霞 《农业生物技术学报》2011,19(6)

本研究通过把构建的短发夹结构RNA(shRNA)真核表达载体导入鸡胚,检测鸡胚性腺分化期质粒载体在鸡胚体内代谢情况及雌性鸡胚性腺中芳香化酶基因(CYP19A1)mRNA表达效率,进而探讨利用该方法在鸡胚体内进行特定基因干涉的可行性.实验针对CYP19A1基因构建了4条特异性表达载体,一条非特异性表达载体.每个实验组选取45个新鲜种蛋作为实验材料进行胚盘下腔注射,并设立空白对照组.鸡胚发育12d时,检测各处理组鸡胚肝脏组织中质粒存在情况,并取其左侧性腺组织进行目标基因的荧光定量分析.研究结果表明,导入组在12日胚龄时所有鸡胚基因组中均可检测到绿色荧光蛋白基因(EGFP);荧光定量结果显示,导入特异性表达载体cyp-580、cyp-1083和cyp-1295后,对应雌性鸡胚性腺CYP19A1 mRNA表达效率显著低于空白对照组,干涉效率分别为:73％、52％和85％;特异性表达载体cyp-1403组CYP19A1mRNA表达效率与空白对照组相比有所降低但无显著性差异.本实验为诱导鸡胚性反转提供了新方法并建立了鸡胚发育期特定基因体内干涉新平台.  相似文献