Expression from two Drosophila promoters in embryos of the migratory locust

Swarna K. Mathi, Virginia K. Walker, Gerard R. Wyatt
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引用次数: 7

Abstract

We have accomplished gene transfer into embryos of Locusta migratoria, the African migratory locust. Freshly oviposited eggs were injected with circular or linear plasmids containing the Drosophila hsp70 promoter and the choramphenicol acetyltransferase (CAT) reporter gene (hsp-cat), or with circular plasmid containing the Drosophila copia promoter fused to CAT (copia-cat). Southern blot analysis showed that the hsp-cat plasmid persisted extrachromosomally for at least 8 days after injection. There was no evidence for plasmid replication. Transient expression from the introduced promoters was determined by monitoring CAT enzyme activity. After injection of hsp-cat, activity was detected at varying levels in 6–8% of day 3 and day 9 embryos. Embryos injected with copia-cat, assayed on day 3, had a greater frequency but no higher level of expression. The described gene transfer system is promising for analysis of other promoters, including those of Locusta.

两个果蝇启动子在飞蝗胚胎中的表达
我们已经完成了将基因转移到非洲迁徙蝗虫的胚胎中。将新鲜的卵注入含有果蝇hsp70启动子和氯霉素乙酰转移酶(CAT)报告基因(hsp-cat)的圆形或线性质粒,或含有与CAT融合的果蝇复制启动子的圆形质粒(复制- CAT)。Southern blot分析显示,注射后hsp-cat质粒在染色体外存活至少8天。没有质粒复制的证据。通过监测CAT酶活性来确定启动子的瞬时表达。注射hsp-cat后,在第3天和第9天的6-8%的胚胎中检测到不同水平的活性。在第3天检测时,注射了复制猫的胚胎表达频率更高,但表达水平没有提高。所描述的基因转移系统有望用于其他启动子的分析,包括Locusta的启动子。
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