DEVELOPMENT OF SPECIFIC RIA AND ELISA TO STUDY TRYPSIN MODULATING OOSTATIC FACTOR IN MOSQUITOS

被引:21
作者
BOROVSKY, D
POWELL, CA
CARLSON, DA
机构
[1] AGR RES & EDUC CTR,FT PIERCE,FL
[2] USDA ARS,VET & MED ENTOMOL LAB,GAINESVILLE,FL 32604
关键词
A-AEGYPTI; PEPTIDE HORMONE; ANTIBODIES; TRYPSIN-LIKE ENZYME; INHIBITION;
D O I
10.1002/arch.940210103
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Trypsin modulating oostatic factor (TMOF), a decapeptide (H-YDPAPPPPPP-OH) that signals the termination of trypsinlike enzyme biosynthesis in the mosquito midgut, was covalently bound to Keyhole Limpet Hemocyanin using N-hydroxy-succinimide and dicyclohexylcarbodiimide. Polyclonal antibodies raised in rabbits against this conjugate were used to develop specific RIA and enzyme linked immunosorbent assay (ELISA) to detect the peptide hormone in female Aedes aegypti. TMOF and its analogs TMOF(B) (H-DYPAPPPPPP-OH), P4 (H-YDPAPPPP-OH), P1 (H-YDPAP-OH), and poly-L-proline were tested with the antiserum. The antiserum fully recognized TMOF and partially recognized P4. Using both RIA and ELISA, we report that the amount of TMOF in the mosquito ovary is 100 +/- 20 ng (S.E.) and 96 +/- 1.4 ng (S.E.), respectively, for each assay. Minute quantities of TMOF a thousandfold lower than in the ovary were found in the mosquito brain, indicating that the hormone is probably not neural but ovarian in origin.
引用
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页码:13 / 21
页数:9
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