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《中国草食动物科学》2016,(3)
根据牙釉质基因在牛X染色体和Y染色体上存在的差异设计巢式引物,对牛静脉血基因组DNA样本以及10枚牛早期胚胎DNA样本进行巢式扩增和电泳分析,以鉴定性别。结果表明:利用此方法能够对牛10 pg量血液基因组DNA进行扩增鉴定性别,雌性产生1条片段长度为311 bp的源于X染色体的条带,雄性产生1条源于X染色体的条带(311 bp)和1条片段长度为251 bp的源于Y染色体的条带,对10枚胚胎进行鉴定,6枚为雄性,4枚为雌性。说明牙釉质基因巢式PCR扩增鉴定牛早期胚胎性别方法可靠,准确性和敏感性较高。 相似文献
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世界上过半数的鸟类为单态性,雌雄难以分辨,对人工饲养繁育造成困难,快速准确地性别鉴定对鸟类繁育研究尤为重要。本文根据已有研究,采用无侵入采样的方式,应用3对引物(2550F/2718R、P2/P8和K7/W1)分别对26种258只鸟类的羽毛样品进行了性别特异性PCR扩增,其中部分产物进一步酶切,经琼脂糖凝胶电泳。结果发现:白鹈鹕雄性为1条321 bp条带,雌性多1条234 bp条带;斑嘴环企鹅雄性为302 bp和50~100 bp条带,雌性多1条367 bp条带;鹤鸵雄性样品为1条约350 bp条带,雌性多1条约150 bp条带。结果表明,羽根作为样品,未经DNA提取直接进行PCR检测,可有效地进行鸟类性别鉴定。 相似文献
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选取牛雄性性别决定基因SRY (sex region of Y chromosome),根据基因序列设计特异引物,应用PCR技术对5头荷斯坦奶牛DNA样品进行扩增,鉴定其性别;并对设计的引物灵敏度进行检测;对已有的公、母各20头荷斯坦奶牛DNA样品进行PCR盲检,获取奶牛高灵敏度特异性引物,用于奶牛性别鉴定。结果表明,4头公牛DNA样品可以扩增出目标条带(66 bp),1头母牛DNA样品无法扩增出条带,阴性对照扩增无条带;最佳引物灵敏度为1.6 pg/μL,可以很好地满足性别鉴定需要。40头个体中,20头个体DNA样品可以扩增出条带,其余20头个体DNA样品无法扩增出条带,检测结果与实际性别对比准确率为100%。试验结果表明,设计的引物灵敏度比较好,能够满足奶牛性别鉴定的需要。 相似文献
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应用Sry-PCR扩增鉴定狍(Capreolus capreolus)的性别 总被引:2,自引:0,他引:2
根据人的SRY基因核心序列设计合成1对引物C1、C2,应用PCR技术对野生狍(Capreolus capreolus)Sry基因(哺乳动物Y染色体DNA雄性特异区)进行扩增.结果在野生狍雄性样本扩增出1条带(221bp).而在雌性样本未见扩增带.显示了Sry基因的性别特异性。应用这1对引物对42个未知性别狍肌肉组织样本进行了性别鉴定.结果雄性22个,雌性20个。对Sry-PCR产物克隆测序,得到185bp的Sry基因部分核苷酸序列。本试验为狍种群性别比率及其种群动态变化机制研究提供了资料。 相似文献
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雏鸽的雌雄外貌形态差别甚微,极难区别。为有效准确地鉴别雏鸽的性别,淘汰非目的性别,从而降低饲养成本,提高经济效益。本研究通过采用PCR方法,选定特殊的性别CHD1基因及鉴定引物,克隆出雏鸽特定性别基因序列鉴定雏鸽的雌雄。结果表明:在雏鸽雌性样本中扩增出1条带(777 bp),而雄性中未见扩增带,显示出CHD1基因的性别特异性,PCR扩增性别鉴定结果与样本解剖后对性腺观察的形态学鉴定结果也完全一致。在87份样品中,阳性克隆结果为25,阴性克隆结果为62,故87只雏鸽中雌鸽25只,雄鸽62只,其准确率达100%。 相似文献
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牙釉质基因鉴定麇鹿性别 总被引:1,自引:0,他引:1
性别鉴定是调查野生种群雌雄性比的重要方法,对野生动物种群管理具有重要意义。而牙釉质(AMEL)基因在性染色体上具有较高的保守性,在鉴定动物性别方面得到了应用,本次试验采用北京麋鹿生态实验中心的15头糜鹿组织样本,其中11个来自雄性麇鹿鹿茸,4个来自雌性糜鹿静脉血液。对所取样本分别进行基因组DNA提取、AMEL基因片段PCR扩增、纯化、测序。得到了AMEL基因鉴定和实际雌雄性别个数差异不显著(P>0.05)。结果表明:雌性麋鹿产生1条322 bpX带和1条N带,雄性麋鹿则产生322 bpX带和277 bpY带以及1条N带,雄鹿(10/10)和雌鹿(4/4)性别鉴定结果分别都与实际性别符合,所以,使用鹿茸角和血液样本进行AMEL.基因扩增电泳分析可以对糜鹿性别鉴定。 相似文献
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Ali Kadivar Habiballah Rashidzadeh Najmeh Davoodian Hasan Nazari Rohallah Dehghani Tafti Heidar Heidari Khoei Hasan Seidi Samani Jahangir Modaresi Ebrahim Ahmadi 《Reproduction in domestic animals》2021,56(2):287-291
Accurate diagnosis of foetal sex in pregnant mare is helpful for many breeders, both for private or commercial purposes. In this study, in order to pre-natal foetal sexing in equine, we used TaqMan duplex real-time PCR to detect the specific regions of SRY and TSPY genes on extracted cell-free foetal DNA from maternal blood. Peripheral blood samples from 50 pregnant Arabian mares with singleton foetuses were collected. Cell-free foetal DNA was extracted from maternal plasma, and duplex real-time PCR assays were performed with TaqMan probes and primers. Amplification of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene was used as control of DNA extraction procedure. From the 50 sampled mares, 28 cases had female and 22 mares had male foetuses. The final results for 46 samples were conclusive, and from them, 43 cases were predicted correctly. Sensitivity, specificity and accuracy of the test were 90.48%, 96% and 93.48%, respectively. In conclusion, a TaqMan duplex real-time PCR was set up to pre-natal detection of foetal sex in equine. The method was fast and decreased the false-positive and false-negative results. The technique can be used as a routine procedure in farms by collecting only a blood sample. 相似文献
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根据兔SRY基因序列设计两对引物作为兔雄性特异性引物,根据兔APP基因序列设计1对引物作为内标引物,分别建立了兔早期胚胎性别鉴定的双重PCR和巢式PCR反应体系,在不同浓度的基因组DNA和兔早期胚胎上进行性别鉴定应用,同时,对兔SRY巢式PCR引物特异性进行了分析。结果表明,多重PCR扩增兔基因组DNA可以准确判定其性别,扩增灵敏度为100pg基因组DNA;多重PCR鉴定24枚兔32细胞桑椹胚性别,只能对整胚成功鉴定。巢式PCR,公兔基因组DNA扩增出282bp的SRY基因片段,母兔没有扩增产物,扩增灵敏度为10pg;对24枚兔32细胞桑椹胚性别鉴定结果表明,巢式PCR可以对少至4个胚胎细胞进行准确鉴定,同一胚胎结果符合率为100%(24/24)。SRY引物只对兔雄性基因组DNA特异,而其他动物(人、牛、绵羊、小鼠)雄性DNA及兔的冲卵液,均无PCR产物。 相似文献
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Mohammad Reza Divar Hassan Sharifiyazdi Mojtaba Kafi 《Veterinary research communications》2012,36(4):215-220
In the current study we aimed to use PCR to investigate the presence of fetal DNA in the bovine (Bos taurus) cervical secretions and maternal serum, and to assess the effectiveness of this method in fetal gender determination. Pregnant uteri and pre-slaughter maternal blood samples were collected from 21 Holstein Frisian cows in a local abattoir. Overall, 13 male and 8 female fetuses were included in the study. Cervical mucus was sampled at the laboratory. After DNA extraction, the PCR amplified a 280?bp fragment from the X-chromosome and a 217?bp fragment from the Y-chromosome based on a sex-related polymorphism in the amelogenin locus. The presence of fetal Y-chromosome was confirmed in seven out of 13 cervical mucus samples collected from cows with male fetuses. Overall test sensitivity for correct sex determination based on PCR assay on cervical samples was equal to 71.4?±?2?%. In contrast, no fetal Y-chromosome DNA was detected in maternal serum samples from cows with male fetuses. This is the first report on validating the presence of fetal DNA material in the bovine cervical mucus and its potential usefulness for fetal sexing. Further investigations are needed to maximize the accuracy and evaluate the practicality of this approach. 相似文献
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本试验采用PCR方法,依据性别基因连锁理论,通过羽毛和血液,不提取鸽子基因组DNA,直接利用特异性引物对其CHD基因的特异性片段进行扩增,琼脂糖凝胶电泳检测,结果显示雄鸽仅有1条带,在700bp左右,雌鸽有2条带,分别在400与700bp左右;利用该检测方法对100羽性别未知的雏鸽进行了性别鉴定,结果为雌鸽48羽、雄鸽52羽,并通过剖解雄鸽的生殖器官进行验证,准确率达到100%。用建立的方法测定了2928羽雏鸽用于蛋鸽留种试验,结果表明,雌鸽为1411羽、雄鸽为1517羽。 相似文献
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In this experiment, the primer pairs were designed, and polymerase chain reaction (PCR) technique to amplify the segments of the partial sex linked gene chromo-helicase-DNA-binding (CHD) to identificate pigeon's sex using feather pulps and blood.We found that two bands were demonstrated in all female egg-laying pigeons, and the sizes of them were 400 and 700 bp, respectively;In all male egg-laying pigeons, only one band appeared with length of 700 bp.100 unknown sex samples were successfully identified (52 males and 48 females).Based on the checking results between molecular electrophoresis of pigeons and the truly sex of maturity pigeons, we detected CHD gene in 2 928 early age pigeons using PCR amplification and electrophoresis in agarose gel.The PCR amplification results showed 1 517 males and 1 411 females. 相似文献