Abstract：
Tissue-specific gene promoters are desired as they provide the specificity needed for control of gene expression in transgenic animals. Here we describe a relatively rapid two-component transient expression assay that was used to identify a gene promoter active in the larval salivary glands of the green blow fly, Lucilia sericata. Sterile L.sericata maggots are widely used for wound debridement. A larval salivary gland gene promoter could be used to make maggots that secrete factors for enhanced wound therapy. Embryos from a line that carry a tetracycline transactivator (tTA)-activated red fluorescent protein gene were injected with plasmid DNA with the tTA gene driven by a constitutive or tissue-specific gene promoter. The hatched larvae were reared on diet and then examined for red fluorescence. A promoter from the LsCG30371 gene was active in the larval salivary glands. The tissue-specificity of the promoter was subsequently confirmed with stable transgenic lines that carried the LsCG30371-tTA gene. The relatively rapid transient expression assay could potentially be used to determine the tissue-specificity of other gene promoters. Further, the stable LsCG30371-tTA lines could be used to make sterile maggots that secrete factors from the salivary glands for enhanced wound healing.
摘要：
组织特异性基因启动子是期望的,因为它们提供控制转基因动物中基因表达所需的特异性。在这里，我们描述了一种相对快速的两组分瞬时表达测定法，该测定法用于鉴定在绿色吹蝇的幼虫唾液腺中有活性的基因启动子，Luciliasericata.无菌的丝黄曲霉广泛用于伤口清创。幼虫唾液腺基因启动子可用于制造分泌因子以增强伤口治疗的the。将携带四环素反式激活因子（tTA）激活的红色荧光蛋白基因的品系的胚胎注入由组成型或组织特异性基因启动子驱动的tTA基因的质粒DNA。在饮食中饲养孵化的幼虫，然后检查红色荧光。来自LsCG30371基因的启动子在幼虫唾液腺中具有活性。随后用携带LsCG30371-tTA基因的稳定转基因株系证实了启动子的组织特异性。相对快速的瞬时表达测定可以潜在地用于确定其他基因启动子的组织特异性。Further,稳定的LsCG30371-tTA细胞系可用于制造无菌mu，从唾液腺分泌因子以增强伤口愈合。
