Production of transgenic quails with high frequency of germ-line transmission using VSV-G pseudotyped retroviral vector.
We report here the production of transgenic quails using a replication-defective pantropic retroviral vector based on Moloney murine leukemia virus (MoMLV) pseudotyped with vesicular stomatitis virus G protein (VSV-G). The retroviral vector was injected into laid quail embryos at the blastodermal stage, and the embryos were incubated to hatch to produce G(0) transgenic quails. Among 134 embryos subjected to viral injection, 37 hatched. The viral vector sequence was detected in the tissues of all G(0) quails. The germ-line transmission efficiency of G(0) quails mated with nontransgenic quails was more than 80% on average. Southern blot analysis revealed that the G(1) transgenic progeny had one to three copies of the transgene. The expression of vector-encoded neomycin-resistance gene under the control of the Rous sarcoma virus (RSV) promoter was observed in several tissues including heart and muscle of both G(1) and G(2) transgenic offspring. Due to the high frequency of germ-line transmission, this method may markedly facilitate the production of transgenic avian.
Mizuarai
S
S
Department of Biotechnology, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Ono
K
K
Yamaguchi
K
K
Nishijima
K
K
Kamihira
M
M
Iijima
S
S
D000818:Animals; D030801:Animals, Genetically Modified / Q000235:genetics*; D015139:Blotting, Southern; D019175:DNA Methylation; D003673:Defective Viruses / Q000235:genetics; D005260:Female; D018628:Gene Dosage; D017930:Genes, Reporter; D005822:Genetic Vectors; D018095:Germ-Line Mutation; D008297:Male; D008562:Membrane Glycoproteins; D008979:Moloney murine leukemia virus / Q000235:genetics; D011784:Quail / Q000235:genetics* / Q000378:metabolism; D012333:RNA, Messenger / Q000096:biosynthesis; D014018:Tissue Distribution; D019076:Transgenes; D014759:Viral Envelope Proteins / Q000235:genetics*
C028391:G protein, vesicular stomatitis virus; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D014759:Viral Envelope Proteins