Cloning and expression of male‐specific protein (MSP) from the hemolymph of greater wax moth, Galleria mellonella L.

Abstract

Male-specific protein (MSP) is a soluble protein that accumulates in high amounts in the hemolymph and other organs of adult male wax moth. The MSP was purified from adult male wax moth by gel filtration and reversed phase column chromatography, and its amino acid sequence was determined. Because of blocked N-terminus, several internal amino acid sequences of MSP were obtained by the in-gel digestion method using trypsin. RT-PCR was conducted using degenerate primers designed from the internal amino acid sequences. 5'-RACE PCR was used to obtain the complete coding region and 5'-UTR sequence. The full length MSP cDNA sequence encodes a 239 amino acid polypeptide with an 18 amino acid signal peptide. The putative mature MSP has a molecular mass of 24,317 Da and an isoelectric point (pI) of 6.00, but shows a molecular mass of 27 kDa on SDS-PAGE. Sequence alignment showed a significant similarity between MSP and juvenile hormone binding proteins (JHBPs) of several lepidopteran species, including G. mellonella.

Cloning and expression analysis of takeout/JHBP family genes of silkworm, Bombyx mori

“…1), suggesting that they are secreted proteins. Most of the previously reported members of this gene family are secreted proteins (Goodman & Chang, 1985; Sarov‐Blat et al ., 2000; Wybrandt & Andersen, 2001; Han et al ., 2003); Manduca JP29 is found in both the nucleus (Palli et al ., 1994) and the insecticyanin granules (which are membrane‐bound) within epidermal cell (Shinoda et al ., 1997), however, and some proteins of this gene family might function in cells. The conserved cysteine residues and hormone binding fragments (Fig.…”

“…mRNA expression of JP29 and Moling of Manduca sexta is development‐dependent and specific to the feeding period of the epidermis (Palli et al ., 1994; Du et al ., 2003). Male‐specific proteins, which have similarity with JHBP, were purified from Galleria and Schistocerca haemolymph (Wybrandt & Andersen, 2001; Han et al ., 2003). The Schistocerca yellow protein binds carotenes and is suggested to have a role in transporting carotenes to the epidermis.…”

Lipophorin uptake by the larval fat body and adult ovary in the wax moth Galleria mellonella

“…Larvae of Galleria mellonella were reared on an artificial diet mixed with a natural diet (bee wax) as described by Han et al. (2003).…”

Molecular Cloning and Characterization of Transferrin from Wax Moth, Galleria mellonella

“…The G. mellonella larvae were reared on an artificial diet mixed with a natural diet (bee wax) as described by Han et al (2003). The insects were kept at 32 1°C and 75 5% relative humidity in darkness.…”