Presnail J K, Hoy M A
Department of Entomology, University of California, Berkeley 94720.
Proc Natl Acad Sci U S A. 1992 Aug 15;89(16):7732-6. doi: 10.1073/pnas.89.16.7732.
A microinjection technique has resulted in stable transformation of the western predatory mite Metaseiulus occidentalis. Early preblastoderm eggs within gravid females were microinjected. The needle was inserted through the cuticle of gravid females into the egg, or the tissue immediately surrounding the egg. This maternal injection method resulted in relatively high levels of survival and transformation. Transformation was achieved without the aid of any transposase-producing helper plasmid. The predatory mite was transformed with a plasmid containing the Escherichia coli beta-galactosidase gene (lacZ) regulated by the Drosophila hsp70 heat-shock promoter. Putatively transformed lines were isolated based on beta-galactosidase activity in first-generation larvae. Transformation was confirmed in the sixth generation by polymerase chain reaction amplification of a region spanning the Drosophila/E. coli sequences. Amplification of a nested region, also spanning the interspecific boundary, provided further evidence for stable transformation. Maternal microinjection may be adaptable to other beneficial arthropods, particularly other phytoseiid mites. Genetic transformation of M. occidentalis may improve its efficiency as a biological control agent as well as provide a method for investigating details of its physiology and ecology.
一种显微注射技术已实现西方捕食螨西方盲走螨（Metaseiulus occidentalis）的稳定转化。对怀孕雌螨体内的早期胚盘前卵进行显微注射。将针头穿过怀孕雌螨的角质层插入卵内，或插入卵周围的组织。这种母体注射方法导致了相对较高的存活率和转化率。在没有任何产生转座酶的辅助质粒的情况下实现了转化。用含有受果蝇hsp70热休克启动子调控的大肠杆菌β-半乳糖苷酶基因（lacZ）的质粒转化捕食螨。根据第一代幼虫中的β-半乳糖苷酶活性分离出假定的转化品系。通过对跨越果蝇/大肠杆菌序列区域的聚合酶链反应扩增，在第六代中证实了转化。对同样跨越种间边界的嵌套区域进行扩增，为稳定转化提供了进一步的证据。母体显微注射可能适用于其他有益节肢动物，特别是其他植绥螨。西方盲走螨的遗传转化可能会提高其作为生物防治剂的效率，并提供一种研究其生理和生态细节的方法。
