Sex determination of bovine preimplantation embryos by oligonucleotide microarray

文献类型： 外文期刊

第一作者： Yang, Hua

作者： Yang, Hua;Zhong, Fagang;Yang, Yonglin;Wang, Xinhua;Liu, Shouren;Zhu, Bin

作者机构：

关键词： Oligonucleotide microarray;Sex determination;SRY gene;Bovine embryos

期刊名称：ANIMAL REPRODUCTION SCIENCE （ 影响因子：2.145； 五年影响因子：2.281 ）

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收录情况： SCI

摘要： The aim has been to set up a rapid and accurate microarray assay using sandwich mode for sex determination of bovine preimplantation embryos. Twelve sequence-specific oligonucleotide capture probes used to discriminate 12 samples were spotted onto thealdehyde-modified glass slides by Arrayer. The 2 recognition probes used to identify coding regions of the sex-determining region of the Y chromosome gene (SRY) and (3-casein (CSN2) reference gene were coupled with biotin. The assay was optimized by using genomic DNA extracted from blood samples of known sex individuals. Polymerase chain reaction (PCR) was used to amplify the fragments in the HMG box region of SRY gene and CSN2 gene with sequence-specific primers. The sex of samples was identified by detecting both the SRY and CSN2 genes simultaneously in 2 reaction cells of microarrays, with the male having SRY and CSN2 signals and the female only CSN2. The sex of 20 bovine preimplantation embryos was determined by oligonucleotide microarray. The protocol was run with a blind test that showed a 100% (82/82) specificity and accuracy in sexing of leukocytes. The bovine embryos were transferred into 20 bovine recipients, with a pregnant rate of 40% (8/20). Three calves were born at term, and 5 fetuseswere miscarried. Their sexes were fully in accordance with the embryonic sex predetermination predicted by oligonucleotide microarray. This suggests that the oligonucleotide microarray method of SRY gene analysis can be used in early sex prediction of bovine embryos in breeding programs.

分类号： Q95

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