Coates C J, Jasinskiene N, Miyashiro L, James A A
Department of Molecular Biology and Biochemistry, Bio Sci II, Room 3205, University of California, Irvine, CA 92697-3900, USA.
Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3748-51. doi: 10.1073/pnas.95.7.3748.
The mariner transposable element is capable of interplasmid transposition in the embryonic soma of the yellow fever mosquito, Aedes aegypti. To determine if this demonstrated mobility could be utilized to genetically transform the mosquito, a modified mariner element marked with a wild-type allele of the Drosophila melanogaster cinnabar gene was microinjected into embryos of a kynurenine hydroxylase-deficient, white-eyed recipient strain. Three of 69 fertile male founders resulting from the microinjected embryos produced families with colored-eyed progeny individuals, a transformation rate of 4%. The transgene-mediated complementation of eye color was observed to segregate in a Mendelian manner, although one insertion segregates with the recessive allele (female-determining) of the sex-determining locus, and a separate insertion is homozygous lethal. Molecular analysis of selected transformed families demonstrated that a single complete copy of the construct had integrated independently in each case and that it had done so in a transposase-mediated manner. The availability of a mariner transformation system greatly enhances our ability to study and manipulate this important vector species.
水手转座元件能够在黄热病蚊子埃及伊蚊的胚胎体细胞中进行质粒间转座。为了确定这种已证明的可移动性是否可用于对蚊子进行基因改造，将一个带有果蝇朱砂基因野生型等位基因标记的改良水手元件显微注射到犬尿氨酸羟化酶缺陷的白眼受体品系的胚胎中。显微注射胚胎产生的69只可育雄性奠基者中有3只产生了有眼色后代个体的家系，转化率为4%。观察到转基因介导的眼色互补以孟德尔方式分离，尽管有一个插入片段与性别决定位点的隐性等位基因（雌性决定）分离，且另一个单独的插入片段是纯合致死的。对选定的转化家系进行分子分析表明，在每种情况下，构建体的单个完整拷贝都以转座酶介导的方式独立整合。水手转化系统的可用性大大增强了我们研究和操纵这种重要病媒物种的能力。
