02160naa a2200229 a 450000100080000000500110000800800410001902400410006010000250010124501230012626000090024952013990025865000220165765000210167965000260170065300190172665300280174570000210177370000170179470000160181177301030182714914082018-05-24       2008    bl uuuu     u00u1 u    #d7 a10.1590/S1678-919920080002000062DOI1 aFIGUEIREDO, J. E. F.  aHeterologous expression of an insecticidal gene from the armed spider (Phoneutria nigriventer).h[electronic resource]  c2008  aInsect-pests are global problems that cause severe damage to crop plants, and their control is commonly based on chemical insecticides. However, negative effects of pesticides on the environment and human health emphasize the necessity to develop alternative methods for insect-pest control. In the present study, a gene coding for the insecticidal peptide TX4(6-1) of the Brazilian armed spider (Phoneutria nígriventer) was cloned in fusion with maltose binding protein (MBP) and expressed in Escherichía colí. The affinity purified protein MBP-GlyTX4 was cleaved with the Xa factor and used for a bioassay against Spodoptera frugíperda and rabbit immunization. Five micrograms GlyTX4 protein injected into the hemocoel of larvae and abdominal cavity of adults produced trembling and uncoordinated movements immediately afier injection and ali adult insects died afier 12h. Afier two days, larvae became paralyzed and the epidermal color changed to dark brown. Furthermore, the development stage was prolonged for two weeks. Altematively, si ices of maize leaves were imbibed with 15 micrograms of the recombinant protein cleaved with the Xa factor and used as diet for larvae. In this experiment, ali larvae died in about 30 minutes. Polyclonal antibodies anti-MBP-GlyTX4 were effective for recognizing MBP and GlyTX4 in whole cell extract from E. coli expressing the recombinant protein.  amolecular cloning  aEscherichia Coli  aSpodoptera Frugiperda  aBioinsecticide  aHeterologous expression1 aKALAPOTHAKIS, E.1 aGOMEZ, M. V.1 aBRESSAN, W.  tJournal of Venomous Animals and Toxins Including Tropical Diseasesgv. 14, n. 2, p. 274-293, 2008.