Ribeiro L A, Mariani P D, Azevedo J L, Rech E L, Schmidt G S, Coutinho L L
Departamento de Produção Animal, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Piracicaba, SP, Brasil.
Braz J Med Biol Res. 2001 Sep;34(9):1115-24. doi: 10.1590/s0100-879x2001000900003.
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.
使用高氦气压生物弹道装置对培养在培养基中的鸡胚进行轰击。为了优化影响转化效率的因素，将受人类巨细胞病毒立即早期增强子/启动子控制的lacZ基因用作报告基因。存活率与转化效率之间存在反比关系。在800磅力/平方英寸的氦气压、500毫米汞柱的真空度、金颗粒以及DNA包被的微粒距胚胎8厘米的飞行距离和胚胎置于距微粒分散锥中心0.5厘米处的条件下，获得了胚胎高存活率和高转化效率的最佳条件。在这些条件下，转化效率为100%，存活率为25%，胚胎体细胞中的表达单位数量在100至1000之间。在最佳条件下轰击的胚胎中也检测到了绿色荧光蛋白的表达。基于所获得的结果，生物弹道过程可被认为是鸡胚转化的一种有效方法，因此可作为研究瞬时基因表达和组织特异性启动子的模型系统。
