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Wang Y.,China Agricultural University | Tong J.,China Agricultural University | Li S.,China Agricultural University | Zhang R.,China Agricultural University | And 8 more authors.
PLoS ONE | Year: 2011

Background: The mammary gland is a conserved site of lipoprotein lipase expression across species and lipoprotein lipase attachment to the luminal surface of mammary gland vascular endothelial cells has been implicated in the direction of circulating triglycerides into milk synthesis during lactation. Principal Findings: Here we report generation of transgenic mice harboring a human lipoprotein lipase gene driven by a mammary gland-specific promoter. Lipoprotein lipase levels in transgenic milk was raised to 0.16 mg/ml, corresponding to an activity of 8772.95 mU/ml. High lipoprotein lipase activity led to a significant reduction of triglyceride concentration in milk, but other components were largely unchanged. Normal pups fed with transgenic milk showed inferior growth performances compared to those fed with normal milk. Conclusion: Our study suggests a possibility to reduce the triglyceride content of cow milk using transgenic technology. © 2011 Wang et al.


Cui D.,China Agricultural University | Zhang L.,China Agricultural University | Li J.,China Agricultural University | Zhao Y.,China Agricultural University | And 4 more authors.
PLoS ONE | Year: 2014

Maternal-fetal IgGs transport occurs either prenatally or postnatally, which confers the newborns with passive immunity before their own immune system has matured. However, little is known about the mechanisms of postnatal IgGs passage in the mammary gland. To investigate how FcRn mediates the IgGs transport in the mammary gland, we first generated bFcRn and anti-HAV mAb transgenic mice, and then obtained HF transgenic mice expressing both transgenes by mating the above two strains. Transgene expression of bFcRn in the four lines was determined by qRT-PCR and western blot. We then localized the expression of bFcRn to the acinar epithelial cells in the mammary gland, and anti-HAV mAb was mainly detected in the acini with weak staining in the acinar epithelial cells. Human IgGs could be detected in both milk and serum of HF transgenic mice by western blot and ELISA. A significantly lower milk to serum ratio of human IgGs in HF mice compared with that of anti-HAV mAb mice, indicating that bFcRn could transport human IgGs across the milk-blood barrier from milk to serum during lactation in HF mice. While, there were no transport of murine IgGs, IgAs, or IgMs. These results provide understandings about the mechanisms of maternal-fetal immunity transfer in the mammary gland. © 2014 Cui et al.


Liu S.,China Agricultural University | Li X.,China Agricultural University | Lu D.,China Agricultural University | Shang S.,China Agricultural University | And 7 more authors.
Transgenic Research | Year: 2012

Transgenesis has been used for expressing human lysozyme (hLZ) in the milk of livestock to improve their disease resistance. Here we describe a human lactoferrin (hLF) BAC as a candidate vector for high-level expression of hLZ in the milk of transgenic mice. Using recombineering, hLF genomic DNA in the hLF BAC was replaced by the hLZ gene (from the ATG start codon to the TAA stop codon), and flanking regions of the hLF gene (a 90-kb 5′ and a 30-kb 3′) were used as transcriptional control elements for hLZ expression. When this construct was used to generate transgenic mice, rhLZ was highly expressed in the milk of four transgenic mouse lines (1. 20-1. 76 g/L), was expressed at a lower level in one additional line (0. 21 g/L). rhLZ from the milk of these transgenic mice exhibited the same antibacterial activity as native hLZ. Our results suggest a potential approach for producing large amounts of hLZ in the milk of livestock. © 2011 Springer Science+Business Media B.V.


He J.,China Agricultural University | Ye J.,China Agricultural University | Li Q.,GenProtein Biotech Ltd. | Feng Y.,Chinese PLA General Hospital | And 7 more authors.
Transgenic Research | Year: 2013

Autosomal dominant polycystic kidney disease (ADPKD) is a common human genetic disease, affecting millions of people worldwide. The progressive growth of cysts in kidneys eventually leads to renal failure in 50 % of patients, and there is currently no effective treatment. Various murine models have been studied to elucidate the disease mechanisms, and much information has been acquired. However, the course of the disease cannot be fully recapitulated using these models. The pig is a suitable model for biomedical research, and pig PKD2 has high similarity to the human ortholog at the molecular level. Here, a mini-pig PKD2 transgenic model was generated, driven by a ubiquitous cytomegalovirus enhancer/promoter. Using somatic cell nuclear transfer, four transgenic pigs with approximately 10 insertion events each were generated. Quantitative real-time PCR and western blotting showed that PKD2 was more highly expressed in transgenic pigs than in wild-type counterparts. Because of the chronic nature of ADPKD, blood urea nitrogen and serum creatinine levels were continuously measured to assess the pig kidney function. The transgenic pigs continue to show no significant alteration in kidney function; it is estimated that 1-2 more years may be required for manifestation of renal cystogenesis in these pigs. © 2013 Springer Science+Business Media Dordrecht.


Tong J.,China Agricultural University | Wei H.,China Agricultural University | Liu X.,China Agricultural University | Hu W.,China Agricultural University | And 4 more authors.
Transgenic Research | Year: 2011

In the swine industry pathogenic infections have a significant negative impact on neonatal survival. Piglets fed with human lysozyme, a natural antibiotic, might be more resistant to gastrointestinal infections. Here we describe the generation of transgenic swine expressing recombinant human lysozyme by somatic cell nuclear transfer. Three cloned female pigs were born, one of which expressed rhLZ at 0.32 ± 0.01 μg/ml in milk, 50-fold higher than that of the pig native lysozyme. Both the transgenic gilts and their progeny appear healthy. Introducing human lysozyme into pigs' milk has a potential to benefit the piglets by enhancing immune function and defending against pathogenic bacteria, thereby increasing the new born survival rate. This advance could be of great value to commercial swine producers. © 2010 Springer Science+Business Media B.V.


Li J.,China Agricultural University | Wei H.X.,China Agricultural University | Li Y.,GenProtein Biotech Ltd. | Li Q.Y.,China Agricultural University | And 2 more authors.
Science China Life Sciences | Year: 2012

To obtain reliable results in quantitative PCR (qPCR) reactions, an endogenous control (EC) gene is needed to correct for systematic variations. In this study, a TaqMan low density array was used to quantify the expression levels of microRNA (miRNA) genes in in vivo fertilized, in vitro fertilized, parthenogenetic and somatic cell nuclear transfer blastocysts. The aim was to identify suitable EC genes for the qPCR analysis of miRNAs in porcine blastocysts. The results showed that thirty-six miRNAs were commonly expressed in the four kinds of embryos and the expression levels of eleven miRNAs were similar in the different embryo types (P-value>0. 05). These 11 miRNAs were selected as candidate EC genes for further analysis and, of these, miR-16 was identified as the most stable EC gene by the GeNorm (a tool based on a pair-wise comparison model that calculates the internal control genes stability measure and determines the most reliable pair of EC genes) and NormFinder (an excel plug-in that uses an ANOVA-based model to estimate intra- and inter-group variation to indicate the single most stable EC gene) programs. In addition, a cell number normalization method validated miR-16 as a suitable EC gene for use in future qPCR analysis of miRNAs in porcine blastocysts. © 2012 Science China Press and Springer-Verlag Berlin Heidelberg.


Tong J.,China Agricultural University | Lillico S.G.,Roslin Institute | Bi M.J.,China Agricultural University | Qing T.,China Agricultural University | And 7 more authors.
Transgenic Research | Year: 2011

One of the primary goals of traditional livestock breeding is to improve growth rate and optimise body size. Growth rate can be significantly increased by integrating a growth hormone (GH) transgene under the control of a ubiquitous promoter, but while such animals do demonstrate increased growth there are also serious deleterious side-effects to the animals health. Here we report the generation and initial characterization of transgenic mice that carried a porcine BAC encoding the porcine GH gene. We show that GH expression is restricted specifically to the pituitary, is associated with elevated IGF-1 levels, and results in growth enhancement. No negative effects to the health of the transgenic animals were detected. This initial characterisation supports the use of BAC pGH transgene in livestock studies. © 2010 Springer Science+Business Media B.V.

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