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

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

  • Tao Gui
  • Meiling Zhang
  • Jianwen Chen
  • Yuanliang Zhang
  • Naru Zhou
  • Zhang, Yu
  • Jia Tao
  • Liucai Sui
  • Yunsheng Li
  • Ya Liu
  • Xiaorong Zhang
  • Yunhai Zhang

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.

OriginalsprogEngelsk
TidsskriftBiotechnology Letters
Vol/bind34
Udgave nummer8
Sider (fra-til)1445-1452
Antal sider8
ISSN0141-5492
DOI
StatusUdgivet - 2012
Eksternt udgivetJa

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