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Activity Detection and Purification of Early Pregnancy Factor in Beef Cattle

Author: HeJiaShuang
Tutor: MaWeiMing
School: Shandong Agricultural University
Course: Clinical Veterinary Medicine
Keywords: Serum of pregnant cows Early Pregnancy Factor Activity Detection Separation and purification
CLC: S823
Type: Master's thesis
Year: 2011
Downloads: 31
Quote: 0
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Abstract


Early Pregnancy Factor (Early Pregnancy Factor, EPF) is present in one of the early pregnancy maternal serum immunosuppressive and growth regulator protein. In 1974 by the Australian scholar Morton, first found in the serum of pregnant rats, can be detected in the mice after 6 h of mating, so named for early pregnancy factor. EPF mammalian fertilization, first appear in the blood having a pregnancy related immune suppression activity of the polypeptide material. Generally agreed that it is a member of the heat shock protein family chaperone protein 10 homologue. Available E rosette inhibition test (E Rosette Inhibition Test, E-RIT) to do biological identification. The early pregnancy secreted EPF can prevent the mother's immune system recognition of the fetus as exogenous substances and the fetus answering. EPF generation depends on an appropriate hormone environment, such as ovarian, fallopian tube, the fertilized egg, placenta, pituitary. EPF in animal fertilization after several hours can be detected in maternal blood, to persist throughout pregnancy. EPF then disappear in the mother after childbirth. EPF can be used for early pregnancy diagnosis, predict early embryonic death and abortion, identification of embryo transfer effects, monitoring the status of the fetus, and secondly, EPF as a gentle, natural immune inhibitor, in the treatment of autoimmune diseases, the prevention of organ transplant rejection reaction in areas such as research value. Therefore, to study this protein production is of great significance to the cattle industry. The experimental collection inseminated 20 h, 25 h and 30 h after the beef cattle serum and vaginal mucus, try to establish a simple and effective E rosette inhibition assay, rapid and accurate detection of early pregnancy factor activity, and determination of blood and vaginal mucus in early pregnancy factor The earliest detection time, in order to determine the earliest opportunity and the best way for early pregnancy detection. The healthy pregnant cow serum collected 1 to 9 months of pregnancy, by the DEAE-52 ion exchange chromatography, Con A-Sepharose 4B affinity chromatography and Heparin Sepharose CL 6B affinity chromatography separation and purification technique, separated from the serum of pregnant cows purified EPF. E rosettes inhibition assay to detect various stages of product EPF activity. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and acrylamide gel electrophoresis and isoelectric focusing analysis of purified EPF isoelectric point. The results showed that, after insemination of beef cattle in between 20h able to detect early pregnancy factor activity, the vaginal mucus detectable time is 25h, pregnant cattle and the nonpregnant bovine serum E rosette inhibition titer was significantly different (P lt; 0.05) purified EPF looped through E rosette inhibition experiments have shown activity and its molecular mass is 53.6 Ku, an isoelectric point of 6.8. Conclusion is ring E rosette inhibition assay early pregnancy factor activity can be identified by the beef cattle pregnancy status, and first insemination 20h within the blood detected purified EPF suppress experimental active E rosettes by SDS-PAGE appears 53.6Ku a molecular weight of protein bands. By polyacrylamide gel isoelectric focusing electrophoresis assay EPF the isoelectric point was 6.8.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Livestock > Cow
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