共查询到17条相似文献,搜索用时 281 毫秒
1.
本研究利用本试验室设计的SRY基因3对常规PCR引物扩增牛SRY基因和羊的SRY基因,结果表明:第2对引物扩增出公牛和公羊基因组的DNA 750 bp左右清晰条带,而母牛、母羊基因组DNA元此扩增条带,说明第2对引物可以应用于牛和羊的早期胚胎性别鉴定; 相似文献
2.
3.
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
4.
依据牛、山羊、兔、猪等哺乳动物SRY基因高度同源性设计一对22 bp的SRY引物,按照3×2×3因子组合,建立了PCR扩增胚胎DNA最适条件。采用此最适条件扩增了15个羊-兔异种克隆胚胎DNA,结果表明PCR法可以用来鉴别哺乳动物胚胎的性别。采用设计的性别鉴定引物,按照最优PCR扩增胚胎DNA条件配制了PCR性别鉴定试剂盒。 相似文献
5.
《中国草食动物科学》2016,(5)
在提取送检的未知肌肉组织样品以及市购的猪肉、牛肉、羊肉样品基因组DNA的基础上,分别选用1对通用引物与3对特异性引物进行扩增,用来检测未知样品的畜种来源。结果显示:通用引物扩增4种样品均出现阳性条带;分别用羊、猪、牛的特异性引物扩增样品DNA,送检未知样品只在牛特异性引物扩增时出现了特异性目的条带。经PCR产物直接测序及同源性比对进行验证确认,该未知样品与牛的同源性为100%,从而验证了该方法的准确性。 相似文献
6.
7.
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
8.
9.
《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
10.
应用PCR技术检测柞蚕微孢子虫 总被引:2,自引:1,他引:1
采用斑迹抽提法提取柞蚕微孢子虫(Nosema pernyi)基因组DNA,基因组DNA的琼脂糖凝胶电泳图谱中有大小约15kb的清晰、完整条带。选用已报道的微粒子属16S rRNA基因的保守序列设计P1/P2和N1/N22对引物,对柞蚕微孢子虫基因组DNA进行PCR扩增,结果2对引物分别扩增出1条大小不同的特异谱条带,其中用引物N1/N2扩增可检测出0.47ng的DNA模板。应用同样的PCR引物、体系和反应条件,可有效扩增出受感染柞蚕幼虫、成虫中的微孢子虫基因组DNA条带。该项检测技术有望应用于柞蚕微粒子病的早期诊断。 相似文献
11.
12.
SRY是Y染色体上具体决定生物雄性性别的基因片段,是多数哺乳动物性别决定基因之一。本试验采用克隆测序SRY基因并结合生物信息学对其序列进行分析,利用软件Codon W分析麦洼牦牛、普通牛、绵羊、山羊、猪、小鼠、鸡和人该基因编码区的密码子偏好性。克隆测序得到SRY基因序列含1个690 bp的开放阅读框,共编码229个氨基酸;其编码区核苷酸序列与普通牛、绵羊、山羊、猪、小鼠、鸡、人相应序列间的一致性分别为99.9%、93.9%、91.2%、76.5%、43.3%、21.4%、69.1%。经聚类分析,麦洼牦牛首先与普通牛聚在一起,绵羊与山羊聚为一类,这两类相聚后再依次与猪、人、小鼠相聚,最后与鸡聚为一类,其结果与以往的生物学分类结果一致。该基因编码的蛋白质分子式为C1155H1827N351O348S11,相对分子质量为2655.0,理论等电点PI为9.55,亲水性平均数为-0.855。其密码子偏好性分析显示,T3s为0.2905、C3s为0.3240、A3s为0.3728、G3s为0.2963,第3位碱基中出现G和C占第3位碱基总量的48%,同义密码子数为61。上述数据表明麦洼牦牛SRY基因编码序列与普通牛、绵羊、山羊、猪、人具有较高的一致性,在物种进化过程中较为保守。该基因对含A的密码子有较高的偏爱性,尽量避开以G结尾的密码子,且其所编码的蛋白质为亲水性蛋白。 相似文献
13.
14.
H.N. Malik D.K. Singhal A. Mukherjee N. Bara S. Kumar S. Saugandhika A.K. Mohanty J.K. Kaushik S. Bag B.C. Das S.K. Bhanja D. Malakar 《Livestock Science》2013,157(1):351-357
The primary objective of this study was to develop a simplified, rapid and authenticated protocol for sexing of caprine embryos. Polymerase chain reaction (PCR) is a powerful tool in preimplantation sex diagnosis, using embryo biopsy at the early developmental stage. Based on the amelogenin gene located on the conserved region of the sex chromosome, a primer pair was used and PCR was established to amplify a 262-bp fragment from the Xchromosome in female goat embryos and 262-bp fragments from the X chromosome and 202-bp fragments from the Y chromosome in male embryos. To validate the reliability of PCR, using the sex-determining region Y (SRY) gene located on the conserved region of Y chromosome, a primer pair was used and PCR was established to amplify a 122-bp fragment specific to the Y chromosome in male embryos. The in vitro-produced goat in vitro fertilisation (IVF)-embryos were made zona free by treating with pronase. The cell number in each embryo was counted before sexing. A single blastomere taken from these embryos was directly used as a template in PCR containing SRY and amelogenin gene-specific primers separately. Of 75 pronase-treated and 60 micromanipulated goat IVF embryos, 33 (44%) and 26 (43.33%) were confirmed as male and 42 (56%) and 34 (56.66%) as female, respectively. The sex-diagnosed embryos were kept in research vitro cleavage (RVCL) medium, and developed into 42.66% and 61.66% morulae and 13.33% and 23.33% blastocysts among pronase-treated and micromanipulated embryos, respectively. The AMELX gene-specific primer served as the internal control and did not interfere with amplification of the Y-specific sequence. In conclusion, a single blastomere sexing protocol based on the SRY and the amelogenin gene is simple, rapid, sensitive and efficient for sex determination in caprine early stage embryos. 相似文献
15.
Hasegaw T Sato F Ishida N Fukushima Y Mukoyama H 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2000,62(10):1109-1110
A quick method for sex determination of horses was developed. Simultaneous amplification of the equine sex-determining region of the Y chromosome gene (SRY) and amelogenin gene (AMEL) accomplished the determination of the presence of both the Y chromosome and SRY gene. In agarose gel electrophoresis, a normal stallion showed 1 SRY band and 3 AMEL (AMELX, AMELY, and AMELX/AMELY heteroduplex) bands, and a normal mare showed a single AMELX band. In XY-mares, 3 AMEL bands were detected as in a normal stallion, but no SRY band. The present method enables a quick diagnosis for XY-mare prior to cytogenetic analysis. 相似文献
16.