JARB Journal of Animal Reproduction and Biotehnology

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eISSN: 2671-4663

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Reproductive & Developmental Biology 2008; 32(4): 255-261

Published online December 30, 2008

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

Rong Xun Han,Su Min Choi,Myung Youn Kim,Yan Shi Quan,Baek-Chul Kim,Yun Fei Diao,Reza Koqani,Chang Sik Park,Dong Il Jin

Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea

Abstract

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0~10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500~600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Keywords: Dairy cattle, Whey proteins, 2‐D gel electrophoresis, Mass spectrometry

Article

Reproductive & Developmental Biology 2008; 32(4): 255-261

Published online December 30, 2008

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Identification of Bovine Pregnancy-Specific Whey Proteins using Two-Dimensional Gel Electrophoresis

Rong Xun Han,Su Min Choi,Myung Youn Kim,Yan Shi Quan,Baek-Chul Kim,Yun Fei Diao,Reza Koqani,Chang Sik Park,Dong Il Jin

Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea,Division of Animal Science & Resources, Research Center for Transgenic and Cloned Pigs, Chungnam National University, Daejeon 338-708, Korea

Abstract

The early diagnosis of bovine pregnancy is an essential component of successful reproductive planning on farms, because lack of bovine pregnancy over the long term results in reproductive failure and low milk yield‐the latter of which is a special concern on dairy farms. This study was designed to identify early pregnancy‐specific whey proteins in bovine, by comparing milk samples collected from cattle during pregnancy (Days 30 and 50) and from non‐pregnant cattle. In this study, differentially expressed proteins in five pregnant and five non‐pregnant Holstein dairy cattle were investigated and compared, using proteomics analysis. The first dimension was applied to a pH 3.0~10.0 strip, by loading a 2‐mg milk protein sample. After the second‐dimension separation was performed, the gels were stained with colloidal Coomassie brilliant blue. The stained gels were scanned and the images were analyzed, to detect variations in protein spots between non‐pregnant and pregnant cattle milk protein spots, using ImageMaster; this was followed by analysis with MALDI TOF‐MS. Analysis of the 2‐DE gel image resulted in a total of approximately 500~600 protein spots, of which 12 spots were differentially expressed, six spots were up‐regulated, and four spots were downregulated; two spots were identified as pregnancy‐specific proteins. These proteins were identified as lactoferrin, NADH dehydrogenase subunit 2, albumin, serum albumin precursor and transferrin. Our results via 2‐D PAGE analysis revealed composite profiles of several milk proteins related to early bovine pregnancy, implying the possible use of these milk proteins in the early detection of bovine pregnancy.

Keywords: Dairy cattle, Whey proteins, 2‐D gel electrophoresis, Mass spectrometry

JARB Journal of Animal Reproduction and Biotehnology

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OPEN ACCESS pISSN: 2671-4639
eISSN: 2671-4663