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1.
性别鉴定技术与人们的日常生活及畜牧业的发展有着密切的关系。精子性别鉴定技术的迅速发展为这一技术的应用提供了广阔的前景。目前最常用的精子性别鉴定技术是流式细胞分选法 ,此法依据 X-精子及 Y-精子 DNA含量的不同将两种精子分开 ,还可以用作分类后精子的检验。因流式细胞分选法存在一些不适于大规模应用的缺陷 ,人们希望能依据分类后的精子之间的对比研究找出特异的蛋白制成抗体 ,设计出高效免疫的精子性别鉴定技术 ,关于这方面的研究已经取得了一定的进展 相似文献
2.
Bovine foetal sex determination—Different DNA extraction and amplification approaches for efficient livestock production
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M Ristanic Lj Stanisic M Maletic U Glavinic V Draskovic N Aleksic Z Stanimirovic 《Reproduction in domestic animals》2018,53(4):947-954
Foetal sex determination using polymerase chain reaction (PCR) in mammals is based on the amplification of gender‐specific foetal DNA sequences circulating in maternal blood. The bovine synepitheliochorial placenta does not allow a direct contact between the trophoblast and the maternal blood, resulting in difficult passage of foetal DNA and, consequently, its very small amounts in maternal bloodstream. Circulating cell‐free foetal DNA (ccffDNA) encompasses short nucleotide fragments (300–600 bp) in maternal circulation. The aim of this study was to assess this non‐invasive method in accurate prenatal sexing in early and late gestational periods in comparison with ultrasound diagnostics. As various DNA isolation and amplification methods were tested, their success in obtaining reliable results was evaluated. Two groups were tested, each consisting of 20 pregnant cows. Blood of a bull and a non‐pregnant heifer was the controls. Extraction of foetal DNA was accomplished by three different methods: using tubes with silicone membranes, a single‐tube extraction without silicone membranes and phenol–chloroform extraction. Following each extraction method, foetal DNA was amplified using PCR and real‐time PCR with both bAML and TSPY primers in a separate reaction. Positive results were obtained only after amplification of foetal DNA extracted with a single‐tube extraction kit. In comparison with ultrasound examination results and foetal gender recorded at birth, the sensitivity of the PCR test was 90% in Group I, but the technique failed to detect male foetuses in Group II. The real‐time PCR test sensitivity in Group I was 90% and in Group II 91.6%. 相似文献
3.
Bijan Soleymani Kamran Mansouri Mohsen Rastegari-Pouyani Shahram Parvaneh Fatemeh Khademi Mehdi Sharifi Tabar Ali Mostafaie 《Reproduction in domestic animals》2021,56(2):270-277
Separation of X and Y chromosome-bearing sperm is an appropriate method for the selection of desired sex of offspring to increase the profit in livestock industries. The purpose of this study was the production of a monoclonal antibody against recombinant bovine sex-determining region Y protein for separation Y sperm. The hybridoma cells from splenocytes of immunized female's balb/C mice and Sp2/0 cells were made. The binding affinity of our monoclonal antibody (mAbSRY2) was compared with mouse monoclonal SRY-15. The Western blot method indicated that mAbSRY2 successfully detected the rbSRY protein. The specificity and sensitivity of mAbSRY2 is comparable to SRY-15 commercially ones. The SRY gene in 100% of bull semen contains the Y chromosome that had the strongest binding affinity to mAbSRY2 was synthesized. In other words, the binding affinity of semen contains the X sperms near the negative control. In general, this immunological method can help to separate X from Y sperms. However, the mAbSRY2 is bind to Y-bearing sexed sperm, but in the future; the sexed sperms need to apply in farms. 相似文献
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利用分离的奶牛X精子冷冻精液生产体内性控胚胎试验 总被引:6,自引:0,他引:6
采用常规精液和分离的X精子冷冻精液对115头超数排卵的青年和经产荷斯坦母牛人工输精生产体内性控胚胎。结果表明,利用2支和3支性控精液给超排青年母牛输精,头均分别获得6.8枚±4.8枚和6.3枚±3.2枚可用胚胎,与常规精液的超排效果(7.2枚±5.2枚)无明显差异;利用2支和3支性控精液给超排经产母牛输精,头均可用胚胎分别为7.5枚±4.5枚和8.6枚±5.6枚,与常规精液的超排效果(6.1枚±3.2枚)相比,分别增加23%(P>0.05)和41%(P>0.05),无显著差异,但解冻后性控精子会明显影响经产母牛的受精率;对用性控精液输精生产的12枚抽样胚胎采用LAMP法进行性别鉴定并移植,结果获得了91.7%的雌性率和36.4%的受胎率。结果提示,X精子的分离过程会在一定程度上影响受精率,用分离的X精子冷冻精液对超排母牛进行人工授精生产体内性控胚胎是可行的。 相似文献
6.
根据绵羊SRY(sex determining region of the Y)基因的723 bp的高度保守序列设计跨度为193 bp的两对巢式雄性特异引物,同时根据绵羊β-B-珠蛋白基因设计跨度为278 bp的两对巢式引物为内标引物,通过析因设计法建立了巢式PCR体系,对公、母羊肝组织基因组进行巢式PCR扩增,经1.5%的琼脂糖电泳检测,公羊同时出现278 bp的β-B-珠蛋白基因扩增带和193 bp的雄性特异扩增带两条带,而母羊只扩增出278 bp常染色体基因序列扩增带.用此巢式PCR体系扩增淋巴细胞,灵敏度≥10 pg DNA(约3个细胞),鉴定全程不超过130 min. 相似文献
7.
白羽鸡快,慢羽纯系及其杂交后代早期生长的研究 总被引:3,自引:0,他引:3
本文通过3个快羽纯系和3个慢羽纯系以及9个自别雌雄商品杂交组合,测定快、慢羽伴性等位基因(k~ 和K)对白羽蛋鸡早期体重的影响。结果,在兼用型鸡中,由同一来源分离出的快羽C系1~10周龄体重明显高于慢羽D系(P<0.05),表现出明显羽速效应;而在蛋用型鸡中,由同一来源分离出的快羽B系早期体重则低于慢羽G系(1~5周龄P<0.05,6~10周龄P<0.10)。表明快慢羽基因的羽速效应因品种(或品系)的不同而异。9个配套组的杂交鸡早期体重均低于双亲均值,明显表现出负的杂交优势,有利于蛋鸡最适体型的筛选。 相似文献
8.
A male specific bovine DNA fragment was used as probe to determine the sex of bovine interphase cells by in situ hybridization. This method also proved useful for determining the sex of bovine embryos. 相似文献
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采用酚-氯仿法和煮沸法从南疆绒山羊血液和早期胚胎中提取基因组DNA,分别以雄羊和雌羊的血液基因组DNA和早期胚胎基因组DNA(约20~30胚胎细胞)为模板,以AMEL引物进行PCR扩增和电泳分析.结果表明:绒山羊5ng量和10pg量血液基因组DNA经扩增后雌性只得到1条非特异性带,雄性得到1条非特异性带和1条特异性带;超微量血液基因组DNA样本(10pg)经巢式扩增和电泳分析能够鉴定绒山羊性别;32枚绒山羊胚胎鉴定结果,雄雌性别比17/15;移植胚胎产羔雄雌比15/14.采用牙釉质基因(AMEL),经巢式扩增和电泳分析能够鉴定绒山羊性别,并对胚胎无损伤;南疆绒山羊早期胚胎性别鉴定结果与胚胎移植后产羔性别结果对比,雌雄性别比率差异不显著(P>0.05). 相似文献