Adamczyk J J, Fescemyer H W, Heckel D G, Gahan L J, Davis R E, Kelly T J
Department of Entomology, Clemson University, SC 29634-0365, USA.
Arch Insect Biochem Physiol. 1996;31(3):237-56. doi: 10.1002/(SICI)1520-6327(1996)31:3<237::AID-ARCH1>3.0.CO;2-V.
Microvitellogenin and vitellogenin cDNA from Manduca sexta (tobacco hornworm) were tested for use as molecular probes to investigate the expression of genes coding for vitellogenins in Spodoptera frugiperda (fall armyworm) and Lymantria dispar (gypsy moth). Cross-hybridization was not observed between the M. sexta cDNAs and S. frugiperda DNA and mRNA. Vitellogenin cDNA from M. sexta did not hybridize to L. dispar DNA or mRNA. However, the 834 bp microvitellogenin cDNA from M. sexta hybridized to an approximately 850 bp transcript in L. dispar mRNA. A 2.5 kb cDNA clone, pz64, was isolated from late last instar larvae of female L. dispar by differential screening. This clone has 38% amino acid sequence (deduced) and 55% nucleic acid sequence similarities with the 3'-end of high molecular weight vitellogenin in Bombyx mori (silkworm). When used as a probe in northern analysis of L. dispar mRNA, this cDNA hybridized to a 5.3 kb transcript in female last instar larvae, pupae, and adults, but not to male last instar larvae and adults. This cDNA did not hybridize to mRNA from M. sexta or S. frugiperda. Expression of the 5.3 kb vitellogenin transcript hybridizing to the 2.5 kb cDNA clone was suppressed in 5-day-old last instar larvae of female L. dispar treated on day 2 with doses of the juvenile hormone analog, methoprene, greater than 10 nmol. Apparently, the high in vivo titer of juvenile hormone during the first 2 days of the last instar represses the transcription of vitellogenin mRNA.
对烟草天蛾的微卵黄原蛋白和卵黄原蛋白cDNA进行了测试,以用作分子探针,研究草地贪夜蛾和舞毒蛾中编码卵黄原蛋白的基因的表达。在烟草天蛾cDNA与草地贪夜蛾DNA和mRNA之间未观察到交叉杂交。烟草天蛾的卵黄原蛋白cDNA与舞毒蛾DNA或mRNA不杂交。然而,烟草天蛾834 bp的微卵黄原蛋白cDNA与舞毒蛾mRNA中一个约850 bp的转录本杂交。通过差异筛选从舞毒蛾末龄幼虫后期的雌性个体中分离出一个2.5 kb的cDNA克隆pz64。该克隆与家蚕中高分子量卵黄原蛋白3'-末端的氨基酸序列(推导)相似度为38%,核酸序列相似度为55%。当用作舞毒蛾mRNA的Northern分析探针时,该cDNA与末龄幼虫后期的雌性、蛹和成虫中的一个5.3 kb转录本杂交,但与末龄幼虫后期的雄性和成虫中的mRNA不杂交。该cDNA与烟草天蛾或草地贪夜蛾的mRNA不杂交。在用剂量大于10 nmol的保幼激素类似物烯虫酯处理第2天的5日龄舞毒蛾末龄幼虫后期雌性个体中,与2.5 kb cDNA克隆杂交的5.3 kb卵黄原蛋白转录本的表达受到抑制。显然,末龄幼虫前两日体内保幼激素的高滴度抑制了卵黄原蛋白mRNA的转录。