Abstract
We generated genetically manipulated chickens and quail by infecting them with a retroviral vector expressing the human growth hormone under the control of chicken ovalbumin promoter/enhancer up to - 3861. bp from the transcriptional start site. The growth hormone was expressed in an oviduct-specific manner and was found in egg white, although its level was low. The DNA sequence of the integrated form of the viral vector in the packaging cells was shown to be truncated and contained only the sequence spanning - 3861 to - 1569. bp. This represented only the DNase I hypersensitive site (DHS) III of the 4 DHSs and lacked the proximal promoter of the ovalbumin control region. We found several TATA-like and other promoter motifs of approximately - 1800. bp and considered that these promoter motifs and DHS III may cause weak but oviduct-specific expression of the growth hormone. To prove this hypothesis and apply this system to oviduct-specific expression of the transgene, the truncated regulatory sequence was fused to an artificial transactivator-promoter system. In this system, initial weak but oviduct-specific expression of the Tet activator from the promoter element in the ovalbumin control sequence triggered a self-amplifying cycle of expression. DsRed was specifically expressed in oviduct cells of genetically manipulated chickens using this system. Furthermore, deletion of a short region possibly containing the promoter elements (- 2112 to - 1569. bp) completely abrogated oviduct-specific expression. Taken together, these results suggest that weak expression of this putative promoter causes oviduct-specific expression of the transgene.
| Original language | English |
|---|---|
| Pages (from-to) | 146-153 |
| Number of pages | 8 |
| Journal | Journal of Bioscience and Bioengineering |
| Volume | 113 |
| Issue number | 2 |
| DOIs | |
| Publication status | Published - Feb 1 2012 |
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All Science Journal Classification (ASJC) codes
- Biotechnology
- Bioengineering
- Applied Microbiology and Biotechnology
Cite this
Chicken oviduct-specific expression of transgene by a hybrid ovalbumin enhancer and the Tet expression system. / Kodama, Daisuke; Nishimiya, Daisuke; Nishijima, Ken ichi; Okino, Yuuki; Inayoshi, Yujin; Kojima, Yasuhiro; Ono, Ken ichiro; Motono, Makoto; Miyake, Katsuhide; Yoshinori, Kawabe; Kyogoku, Kenji; Yamashita, Takashi; Kamihira, Masamichi; Iijima, Shinji.
In: Journal of Bioscience and Bioengineering, Vol. 113, No. 2, 01.02.2012, p. 146-153.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Chicken oviduct-specific expression of transgene by a hybrid ovalbumin enhancer and the Tet expression system
AU - Kodama, Daisuke
AU - Nishimiya, Daisuke
AU - Nishijima, Ken ichi
AU - Okino, Yuuki
AU - Inayoshi, Yujin
AU - Kojima, Yasuhiro
AU - Ono, Ken ichiro
AU - Motono, Makoto
AU - Miyake, Katsuhide
AU - Yoshinori, Kawabe
AU - Kyogoku, Kenji
AU - Yamashita, Takashi
AU - Kamihira, Masamichi
AU - Iijima, Shinji
PY - 2012/2/1
Y1 - 2012/2/1
N2 - We generated genetically manipulated chickens and quail by infecting them with a retroviral vector expressing the human growth hormone under the control of chicken ovalbumin promoter/enhancer up to - 3861. bp from the transcriptional start site. The growth hormone was expressed in an oviduct-specific manner and was found in egg white, although its level was low. The DNA sequence of the integrated form of the viral vector in the packaging cells was shown to be truncated and contained only the sequence spanning - 3861 to - 1569. bp. This represented only the DNase I hypersensitive site (DHS) III of the 4 DHSs and lacked the proximal promoter of the ovalbumin control region. We found several TATA-like and other promoter motifs of approximately - 1800. bp and considered that these promoter motifs and DHS III may cause weak but oviduct-specific expression of the growth hormone. To prove this hypothesis and apply this system to oviduct-specific expression of the transgene, the truncated regulatory sequence was fused to an artificial transactivator-promoter system. In this system, initial weak but oviduct-specific expression of the Tet activator from the promoter element in the ovalbumin control sequence triggered a self-amplifying cycle of expression. DsRed was specifically expressed in oviduct cells of genetically manipulated chickens using this system. Furthermore, deletion of a short region possibly containing the promoter elements (- 2112 to - 1569. bp) completely abrogated oviduct-specific expression. Taken together, these results suggest that weak expression of this putative promoter causes oviduct-specific expression of the transgene.
AB - We generated genetically manipulated chickens and quail by infecting them with a retroviral vector expressing the human growth hormone under the control of chicken ovalbumin promoter/enhancer up to - 3861. bp from the transcriptional start site. The growth hormone was expressed in an oviduct-specific manner and was found in egg white, although its level was low. The DNA sequence of the integrated form of the viral vector in the packaging cells was shown to be truncated and contained only the sequence spanning - 3861 to - 1569. bp. This represented only the DNase I hypersensitive site (DHS) III of the 4 DHSs and lacked the proximal promoter of the ovalbumin control region. We found several TATA-like and other promoter motifs of approximately - 1800. bp and considered that these promoter motifs and DHS III may cause weak but oviduct-specific expression of the growth hormone. To prove this hypothesis and apply this system to oviduct-specific expression of the transgene, the truncated regulatory sequence was fused to an artificial transactivator-promoter system. In this system, initial weak but oviduct-specific expression of the Tet activator from the promoter element in the ovalbumin control sequence triggered a self-amplifying cycle of expression. DsRed was specifically expressed in oviduct cells of genetically manipulated chickens using this system. Furthermore, deletion of a short region possibly containing the promoter elements (- 2112 to - 1569. bp) completely abrogated oviduct-specific expression. Taken together, these results suggest that weak expression of this putative promoter causes oviduct-specific expression of the transgene.
UR - http://www.scopus.com/inward/record.url?scp=84856799122&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84856799122&partnerID=8YFLogxK
U2 - 10.1016/j.jbiosc.2011.10.006
DO - 10.1016/j.jbiosc.2011.10.006
M3 - Article
VL - 113
SP - 146
EP - 153
JO - Journal of Bioscience and Bioengineering
JF - Journal of Bioscience and Bioengineering
SN - 1389-1723
IS - 2
ER -