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Functional analysis of bovine Nramp1 and production of transgenic cloned embryos in vitro

Published online by Cambridge University Press:  17 May 2013

Xiang Cheng ,
Xiaoli Yu ,
Yajun Liu ,
Jie Deng ,
Xiaoling Ma  and
Huayan Wang
Xiang Cheng
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Xiaoli Yu
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Yajun Liu
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Jie Deng
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Xiaoling Ma
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
Huayan Wang*
Affiliation:
Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China.
*
All correspondence to: Huayan Wang. Department of Animal Biotechnology, College of Veterinary Medicine, Northwest A&F University, Yangling, Shaanxi 712100, China. Tel: +86 029 87080069. Fax: +86 029 87080068. E-mail: hhwang101@163.com
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Summary

Natural resistance-associated macrophage protein 1 (Nramp1) plays an important role in restraining the growth of intracellular pathogens within macrophages. In this study, Nramp1 cDNA was cloned from Qinchuan cattle and its anti-bacterial activity was demonstrated as being able to significantly inhibit the growth of Salmonella abortusovis and Brucella abortus in macrophages. Calf fibroblasts stably transfected with pSP–NRAMP1–HA vector were used to reconstruct bovine embryos by somatic cell nuclear transfer (SCNT). Reconstructed embryos were maturated in vitro and the blastocyst formation rate (14.0%) was similar to that of control embryos (14.5%). Transgenic blastocysts were transplanted into 43 recipient cattle, of which 14 recipients became pregnant as evidenced by non-return estrus and by rectal palpation. One fetus was aborted after 6½ months of pregnancy and transgene integration was confirmed by semi-quantitative polymerase chain reaction. Together, this study showed that bovine Nramp1 retains biological function against the growth of intracellular bacteria and can be used to reconstruct embryos and produce Nramp1 transgenic cattle, which may benefit the animal and enhance their ability to prevent attack by intracellular pathogens.

Keywords

Antibacterial activityBovineNramp1Reconstructed embryoTransgenic animal
Type
Research Article
Information
Zygote , Volume 23 , Issue 1 , February 2015 , pp. 83 - 92
Copyright
Copyright © Cambridge University Press 2013 

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