Molecular weight estimation of esterase isoenzymes in closely related Drosophila Species (Diptera: Drosophilidae) in non-denaturing polyacrylamide gel electrophoresis

Abstract

A method that allows the measure of molecular weight of two well-known and closely related esterases fromDrosophila mojavensis and its sibling species, D. arizonae, is here described, using native polyacrylamide gel electrophoresis at several concentrations, applying Fergunson´s principles. These enzymes, namely presented

Gene Duplication and Subsequent Differentiation of Esterases in Cactophilic Drosophila Species

“…The results presented there are part of this study. Therefore, in this study, we present the results that were not shown in Mateus et al (2009) mulleri, which could be a result of differences in the I.P. values of theses enzymes (Table 3).…”

“…The set of proteins known as esterases constitute one of the most heavily studied groups of isozymes. In the Drosophila mulleri complex, which is the subject of this study, esterases have been extensively studied in several species, including D. serido (Lapenta et al, 1995(Lapenta et al, , 1998, D. buzzatii (East, 1982;Barker, 1994;Lapenta et al, 1995Lapenta et al, , 1998, D. mojavensis (Zouros et al, 1982;Zouros & Van Delden, 1982;Pen et al, 1984Pen et al, , 1986aPen et al, , 1986bMateus et al, 2009), D. arizonae (Zouros et al, 1982;Ceron, 1988;Mateus et al, 2009), D. aldrichi (F. M. Johnson et al, 1968Kambysellis et al, 1968) and D. mulleri (F. M. Johnson et al, 1968;Kambysellis et al, 1968;Richardson & Smouse, 1976;Ceron, 1988). Zouros et al (1982) detected two esterases with different patterns of temporal and tissuespecific expression in Drosophila mojavensis and D. arizonae (formerly D. arizonensis).…”

“…The MW estimation was performed using the method adapted by Mateus et al (2009) from Hedrick and Smith (1968). The following standard MW proteins were used: myoglobin (17.8 kD), soybean trypsin inhibitor (24 kD), carbonic anhydrase (29 kD), ovalbumin (45 kD), human serum albumin (66 kD) and phosphorylase-b (97.4 kD).…”

“…To determine the MW of both enzymes in all six Drosophila species analyzed, the technique described by Mateus et al (2009) was applied using 6% to 12% PAGE and the same MW markers. The results presented there are part of this study.…”

“…Johnson, 1973Johnson, , 1974, and gene duplication has been used as one explanation for their evolution (Zouros et al, 1982;Pen et al, 1990;Mateus et al, 2009). Several studies on the changes in esterase activity during development in species of the D. repleta group have detected two main β-esterases that show different tissue-specific and temporal expression patterns (Zouros et al, 1982;East, 1982;Pen et al, 1984;Pen et al, 1986aPen et al, , 1986bPen et al, 1990;Mateus et al, 2009). One esterase, named EST-4, is present only in later third instar larvae and early pupae and has a high concentration in the carcass.…”