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| The Gene Cloning and Blood Concentration Analysis of EPF After Artificial Insemination in Yak (Bos grunniens) |
| MA Jin-Biao1, FAN Jiang-Feng1,2,*, YU Si-Jiu2, CUI Yan2, HAN Xiao-Hong2, MA Yue1, HE Hong-Hong2, LI Qin2, ZHAO Sheng-Xian1, ZHANG Xing-Yun1, CAO Yan-Tao1, Bao Mei1 |
1 College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China;
2 Technology and Research Center of Gansu Province for Embryonic Engineering of Bovine and Sheep & Goat, Lanzhou 730070, China |
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Abstract Early Pregnancy Factor (EPF) is a bioactive protein that firstly separated from gestational mammal serum. EPF prevents maternal rejection of the fetus in the manner of regulating immunosuppressive action. To analyze the biological characteristics and variation pattern of EPF in yak (Bos grunniens) blood, verify the correlation between blood EPF concentration and pregnancy, yak ovarian EPF gene sequencing was conducted after RT-PCR. The physicochemical properties and protein structure of yak EPF were predicted using bioinformatics related software. Blood samples were collected from 69 yaks during 20~40 d after artificial insemination (AI). The EPF level was measured by enzyme-linked immuno sorbent assay (ELISA). The complete coding sequence (CDS) of yak EPF gene was cloned and submitted to GenBank (No. MK992914). The open reading frame was 207 bp in length and encoded 69 amino acids. The ELISA results showed that the serum EPF level was closely related to the pregnancy status of female yak. In pregnancy group, the EPF concentration in serum gradually increased after the 20 d of AI, and reached the highest value on the 28 d and subsequently decreased slightly. In the non-pregnant group, the EPF concentration in blood maintained a stable and low concentration during 20~40 d after AI, and was significantly lower than that in pregnancy group (P<0.01). These results implied that blood EPF, as a candidate factor, could be used in early pregnancy diagnosis. It shortened the confirming time to 20 days by using blood EPF inspection in pregnancy diagnosis of yak. This study provides basic data for the research and development of products related to early pregnancy diagnosis of yaks and the establishment of technical systems.
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Received: 11 October 2019
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Corresponding Authors:
*, fanjf@gsau.edu.cn
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