Consistent production of transgenic chickens using replication-deficient retroviral vectors and high-throughput screening procedures.
We have developed a novel method of DNA extraction combined with a high-throughput method of gene detection allowing thousands of potentially transgenic chicks to be screened quickly and reliably. By using this method and a replication-deficient retroviral vector based on avian leukosis virus (ALV), we have demonstrated germline transmission of three different transgenes. Several generations of chickens carrying intact transgenes were produced, validating the use of the ALV retroviral vectors for large-scale production of transgenic flocks. Fourth-generation chicks that were nontransgenic, hemizygous, or homozygous for the transgene were identified with the combined genetic screening methods.
Harvey
A J
AJ
AviGenics, Inc., Georgia BioBusiness Center, Athens, Georgia 30605, USA. harvey@avigenics.com
Speksnijder
G
G
Baugh
L R
LR
Morris
J A
JA
Ivarie
R
R
D000483:Alleles; D000818:Animals; D030801:Animals, Genetically Modified; D015139:Blotting, Southern; D002645:Chickens / Q000235:genetics*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004247:DNA / Q000097:blood; D015247:Deoxyribonuclease HindIII; D004351:Drug Resistance / Q000235:genetics; D005822:Genetic Vectors; D006720:Homozygote; D008297:Male; D009355:Neomycin; D010047:Ovalbumin / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic; D012190:Retroviridae / Q000235:genetics*; D019076:Transgenes / Q000235:genetics; D014779:Virus Replication
D010047:Ovalbumin; D004247:DNA; D015500:Chloramphenicol O-Acetyltransferase; D015247:Deoxyribonuclease HindIII; D009355:Neomycin