In vitro evaluation of a mammary gland specific expression vector encoding recombinant human lysozyme for development of transgenic dairy goat embryos

TY - JOUR

T1 - In vitro evaluation of a mammary gland specific expression vector encoding recombinant human lysozyme for development of transgenic dairy goat embryos

AU - Gui, Tao

AU - Zhang, Meiling

AU - Chen, Jianwen

AU - Zhang, Yuanliang

AU - Zhou, Naru

AU - Zhang, Yu

AU - Tao, Jia

AU - Sui, Liucai

AU - Li, Yunsheng

AU - Liu, Ya

AU - Zhang, Xiaorong

AU - Zhang, Yunhai

PY - 2012/8

Y1 - 2012/8

N2 - A vector expressing human lysozyme (pBC1-hLYZ-GFP-Neo) was evaluated for gene and protein expression following liposome-mediated transformation of C-127 mouse mammary cancer cells. Cultures of G418-resistant clones were harvested 24-72 h after induction with prolactin, insulin and hydrocortisone. Target gene expression was analyzed by RT-PCR and Western blot and recombinant human lysozyme (rhLYZ) bacteriostatic activity was also evaluated. The hLYZ gene was correctly transcribed and translated in C-127 cells and hLYZ inhibited gram-positive bacterial growth, indicating the potential of this expression vector for development of a mammary gland bioreactor in goats. Guanzhong dairy goat skin fibroblasts transfected with pBC1-hLYZ-GFP-Neo were used to construct a goat embryo transgenically expressing rhLYZ by somatic nuclear transplantation with a blastocyst rate of 9.0 ± 2.8 %. These data establish the basis for cultivation of mastitis-resistant hLYZ transgenic goats.

AB - A vector expressing human lysozyme (pBC1-hLYZ-GFP-Neo) was evaluated for gene and protein expression following liposome-mediated transformation of C-127 mouse mammary cancer cells. Cultures of G418-resistant clones were harvested 24-72 h after induction with prolactin, insulin and hydrocortisone. Target gene expression was analyzed by RT-PCR and Western blot and recombinant human lysozyme (rhLYZ) bacteriostatic activity was also evaluated. The hLYZ gene was correctly transcribed and translated in C-127 cells and hLYZ inhibited gram-positive bacterial growth, indicating the potential of this expression vector for development of a mammary gland bioreactor in goats. Guanzhong dairy goat skin fibroblasts transfected with pBC1-hLYZ-GFP-Neo were used to construct a goat embryo transgenically expressing rhLYZ by somatic nuclear transplantation with a blastocyst rate of 9.0 ± 2.8 %. These data establish the basis for cultivation of mastitis-resistant hLYZ transgenic goats.

KW - Animals

KW - Animals, Genetically Modified

KW - Bioreactors

KW - Cell Line, Tumor

KW - Embryo, Mammalian

KW - Female

KW - Genetic Vectors

KW - Goats

KW - Green Fluorescent Proteins

KW - Humans

KW - Male

KW - Mammary Glands, Animal

KW - Mice

KW - Muramidase

KW - Polymerase Chain Reaction

KW - Recombinant Proteins

U2 - 10.1007/s10529-012-0930-7

DO - 10.1007/s10529-012-0930-7

M3 - Journal article

C2 - 22526423

SN - 0141-5492

VL - 34

SP - 1445

EP - 1452

JO - Biotechnology Letters

JF - Biotechnology Letters

IS - 8

ER -