Title: A biolistic process for in vitro gene transfer into chicken embryos

Author(s):  Ribeiro LA, Mariani PDSC, Azevedo JL, Rech EL, Schmidt GS, Coutinho LL

Source:  BRAZILIAN JOURNAL OF MEDICAL AND BIOLOGICAL RESEARCH 34 (9): 1115-1124 SEP 2001

Document Type:  Article

Language:  English

Abstract:  Chicken embryos kept in culture medium were bombarded using a high helium gas pressure biolistic device. To optimize the factors that affect transformation efficiency, the lacZ gene under control of the human cytomegalovirus immediate early enhancer/promoter was used as a reporter gene. There was an inverse relationship between survival rate and transformation efficiency. The best conditions obtained for high embryo survival and high transformation efficiency were achieved with 800 psi helium gas pressure, 500 mmHg vacuum, gold particles, an 8 cm DNA-coated microparticle flying distance to the embryo and embryo placement 0.5 cm from the center of the particle dispersion cone. Under these conditions, transformation efficiency was 100%, survival rate 25% and the number of expression units in the embryo body cells ranged from 100 to 1,000. Expression of green fluorescent protein was also detected in embryos bombarded under optimal conditions. Based on the results obtained, the biolistic process can be considered an efficient method for the transformation of chicken embryos and therefore can be used as a model system to study transient gene expression and tissue-specific promoters. Author Keywords:  biolistic process; chicken embryo; gene transfer; beta-galactosidase; green fluorescent protein; GFP

KeyWords Plus:  HIGH-VELOCITY MICROPROJECTILES; PRIMORDIAL GERM-CELLS; PARTICLE BOMBARDMENT; FOREIGN GENES; IN-VIVO; BALLISTIC TRANSFECTION; TRANSIENT EXPRESSION; TRANSGENIC BIRDS; DNA; TRANSFORMATION

Addresses:  Coutinho LL (reprint author), USP, ESALQ, Dept Anim Prod, Av Padua Dias 11, Piracicaba, SP BR-13418900 Brazil
USP, ESALQ, Dept Anim Prod, Piracicaba, SP BR-13418900 Brazil
Univ Sao Paulo, Escola Super Agr Luiz de Queiroz, Dept Genet, Piracicaba, SP BR-13418900 Brazil
EMBRAPA, CENARGEN, SAIN Parque Rural, Brasilia, DF Brazil
EMBRAPA Suinos & Aves, Concordia, SC Brazil

Publisher:  ASSOC BRAS DIVULG CIENTIFICA, FACULDADE MEDICINA, SALA 21, 14049 RIBEIRAO PRETO, SAO PAULO, BRAZIL

Subject Category:  MEDICINE, RESEARCH & EXPERIMENTAL; BIOLOGY

IDS Number:  474QD

ISSN:  0100-879X