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Journal of Zhejiang University SCIENCE B 2005 Vol.6 No.2 P.137~141

http://doi.org/10.1631/jzus.2005.B0137


Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo


Author(s):  GAO Bo, SUN Huai-chang, SONG Cheng-yi, WANG Zhi-yue, CHEN Qin, SONG Hong-qin

Affiliation(s):  School of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China; more

Corresponding email(s):   gaoboyzu@hotmail.com, sun78564@hotmail.com

Key Words:  Chicken ovalbumin gene regulatory regions, Oviduct-specific expression vector, Oviduct epithelium, In vivo expression


GAO Bo, SUN Huai-chang, SONG Cheng-yi, WANG Zhi-yue, CHEN Qin, SONG Hong-qin. Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo[J]. Journal of Zhejiang University Science B, 2005, 6(2): 137~141.

@article{title="Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo",
author="GAO Bo, SUN Huai-chang, SONG Cheng-yi, WANG Zhi-yue, CHEN Qin, SONG Hong-qin",
journal="Journal of Zhejiang University Science B",
volume="6",
number="2",
pages="137~141",
year="2005",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2005.B0137"
}

%0 Journal Article
%T Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo
%A GAO Bo
%A SUN Huai-chang
%A SONG Cheng-yi
%A WANG Zhi-yue
%A CHEN Qin
%A SONG Hong-qin
%J Journal of Zhejiang University SCIENCE B
%V 6
%N 2
%P 137~141
%@ 1673-1581
%D 2005
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2005.B0137

TY - JOUR
T1 - Transfection and expression of exogenous gene in laying hens oviduct in vitro and in vivo
A1 - GAO Bo
A1 - SUN Huai-chang
A1 - SONG Cheng-yi
A1 - WANG Zhi-yue
A1 - CHEN Qin
A1 - SONG Hong-qin
J0 - Journal of Zhejiang University Science B
VL - 6
IS - 2
SP - 137
EP - 141
%@ 1673-1581
Y1 - 2005
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2005.B0137


Abstract: 
To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5′-flanking sequence and 3.0 kb of the 3′-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5′-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the b-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.

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Reference

[1] Bruno, L., Gunther, S., 1989. Cell-type specificity of regulatory elements identified by linker scanning mutagenesis in the promoter of the chicken lysozyme gene. Nucleic Acids Research, 17(21):8451-8463.

[2] Haecker, S.A., Muramatsu, T., Sensenbaugh, K.R., Sanders, M.M., 1995. Repression of the ovalbumin gene involves multiple negative elements including a ubiquitous transcriptional silencer. Mol Endocrinol, 9(9):1113-1126.

[3] Hiroshi, O., Hyi-Man, P., Akihiro, N., Ryuzo, S., Jun-Ichi, O., Tatsuo, M., 1998. Synthesis of human erythropoietin in vivo in the oviduct of laying hens by localized in vivo gene transfer using electroporation. Poultry Science, 77:299-302.

[4] Hiroshi, T., Hisako, W., Yasushige, O., Hyi-Man, P., Tatsuo, M., 2002. Human alkaline phosphatase expression and secretion into chicken eggs in vivo gene electroporration in the oviduct of laying hens. Biochemical and Biophysical Research Communications, 292:88-93.

[5] Liu, Y.B., Yu, M.Z., Shen, X.Z., 2001. The establishment of a temporary expression system in chicken oviduct epithelium. Developmental & Reproductive Biology, 10(1):13-19.

[6] Masami, Y., Takami, O., 1990. Transfection of β-casein chimeric gene and hormonal induction o its expression in primary murine mammary epithelial cells. Proc Natl Acad Sci USA, 87:3670-3674.

[7] Muramatsu, T., Hiramatsu, H., Okumura, J., 1995. Induction of ovalbumin mRNA by ascorbic acid in primary cultures of tubular gland cells of the chicken oviduct. Biochemistry and Molecular Biology, 112:2209-2216.

[8] Park, H.M., Okumura, J., Muramatsu, T., 1995. Modulation of transcriptional activity of the chicken ovalbumin gene promoter in primary cultures of chicken oviduct cell: effects of putative regulatory element the 5(-flanking region. Biochem Mol Biol Int, 36(4):811-816.

[9] Park, H.M., Muramatsu, T., 1999. In vivo manipulation of foreign gene expression by steroid administeration in the oviduct of laying hens. Journal of Endocrinology, 163:173-179.

[10] Sanders, M.M., McKnight, G.S., 1988. Positive and negative regulatory elements control the steroid-responsive ovalbumin promoter. Biochemistry, 27:6550-6557.

[11] Yu, L., Zhao, J., Zhang, Y.L., 2001. Construction of the expressing vector of 5(-flanking regulatory regions o the chicken ovalbumin gene and its transient expression in chicken primary oviduct cell and chicken fibroblasts cell cultures. Chinese Journal of Veterinary Science, 21(1):21-24 (in Chinese).

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