title: Regulation of sheep α-TTP by dietary vitamin E and preparation of monoclonal antibody for sheep α-TTP.
abstract: α-Tocopherol transfer protein (α-TTP) is a cytosolic protein that plays an important role in regulating concentrations of plasma α-tocopherol (the most bio-active form of vitamin E). Despite the central roles that α-TTP plays in maintaining vitamin E adequacy, we have only recently proved the existence of the α-TTP gene in sheep and, for the first time, cloned its full-length cDNA. However, the study of sheep α-TTP is still in its infancy. In the present study, thirty-five local male lambs of Tan sheep with similar initial body weight were randomly divided into five groups and fed with diets supplemented with 0 (control group), 20, 100, 200, 2000IU·sheep(-1)·d(-1) vitamin E for 120 days. At the end of the experiment, the plasma and liver vitamin E contents were analyzed first and then α-TTP mRNA and protein expression levels were determined by quantitative real-time PCR (qRT-PCR) and Western-blot analysis, respectively. In addition, as no sheep α-TTP antibody was available, a specific monoclonal antibody (McAb) against the ovine α-TTP protein was prepared. The effect of vitamin E supplementation was confirmed by the significant changes in the concentrations of vitamin E in the plasma and liver. As shown by qRT-PCR and Western-blot analysis, dietary vitamin E does not affect sheep α-TTP gene expression, except for high levels of vitamin E supplementation, which significantly increased expression at the protein level. Importantly, the specific sheep anti-α-TTP McAb we generated could provide optimal recognition in ELISA, Western-blot and immunohistochemistry assays, which will be a powerful tool in future studies of the biological functions of sheep α-TTP.
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Kun
authors.initials: K
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China. Electronic address: luohailing@cau.edu.cn.
authors.forename: Hai-Ling
authors.initials: HL
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Zhao-Yun
authors.initials: ZY
authors.lastname: Zuo
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Hui-Na
authors.initials: HN
authors.lastname: Jia
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Yu-Wei
authors.initials: YW
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Yan-Fei
authors.initials: YF
authors.lastname: Chang
authors.affiliation: State Key Laboratory of Animal Nutrition, College of Animal Science and Technology, China Agricultural University, Beijing 100193, PR China.
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Jiao
mesh_terms: D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000737:chemistry; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D019587:Dietary Supplements; D005260:Female; D015870:Gene Expression; D005786:Gene Expression Regulation; D022781:Hepatocytes / Q000378:metabolism; D006825:Hybridomas; D007150:Immunohistochemistry; D008099:Liver / Q000166:cytology / Q000378:metabolism; D008297:Male; D008807:Mice, Inbred BALB C; D034561:Sheep, Domestic / Q000235:genetics / Q000378:metabolism*; D014810:Vitamin E / Q000008:administration & dosage / Q000502:physiology*
chemical_list: D058846:Antibodies, Monoclonal, Murine-Derived; D002352:Carrier Proteins; C017763:alpha-tocopherol transfer protein; D014810:Vitamin E
keywords: Expression pattern; Monoclonal antibody; Tan sheep; Vitamin E status; α-Tocopherol transfer protein