Selection of in vitro produced, transgenic embryos by nested PCR for efficient production of transgenic goats.
The production of valuable pharmaceutical proteins using transgenic animals as bioreactors has become one of the goals of biotechnology. However, the efficiency of producing transgenic animals by means of pronuclear microinjection is low. This may be attributed in part to the low integration rate of foreign DNA. Therefore, a large number of recipients are required to produce transgenic animals. We recently developed a transgenic procedure that combined the techniques of goat oocyte in vitro maturation (IVM), in vitro fertilization (IVF), microinjection, preimplantation selection of the transgenic embryos with nested PCR and transferring the transgenic embryos into the recipient goat uterus to produce transgenic goats. Thirty-seven transgenic embryos determined by nested PCR were transferred to thirty-two recipient goats. In the end, four live-born kids were produced. As predicted, all the live kids were transgenic as identified by PCR as well as Southern blot hybridization, The integration rate was 100% (4/4) which was completely in accordance with the results of embryo preimplantation detection. The results showed a significant decrease in the number of recipients required as only 8 recipients (32/4) were needed to obtain one live transgenic goat. We suggest that the transgenic system described herein may provide an improved way to efficiently produce transgenic goats on a large scale.
Huang
S Z
SZ
Shanghai Institute of Medical Genetics, Shanghai Children's Hospital, PR China.
Huang
Y
Y
Chen
M J
MJ
Zeng
F Y
FY
Ren
Z R
ZR
Zeng
Y T
YT
D000818:Animals; D030801:Animals, Genetically Modified / Q000196:embryology* / Q000235:genetics / Q000502:physiology; D000831:Animals, Newborn; D004247:DNA / Q000737:chemistry / Q000235:genetics; D004624:Embryo Transfer / Q000662:veterinary*; D005164:Factor IX / Q000096:biosynthesis; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D006041:Goats / Q000196:embryology* / Q000235:genetics / Q000502:physiology; D008297:Male; D009865:Oocytes / Q000502:physiology; D016133:Polymerase Chain Reaction; D011247:Pregnancy
D005164:Factor IX; D004247:DNA