Cloning and expression of male-specific protein (MSP) from the hemolymph of greater wax moth, Galleria mellonella L.
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.
Han
Jikhyon
J
Laboratory of Animal Macromolecules Physiology and Biochemistry, School of Life Sciences and Biotechnology, Korea University, Seoul, Korea.
Lee
Chang Seok
CS
Yun
Chi Young
CY
Lee
Bong Hee
BH
Ko
Young Gyu
YG
Kang
Chang Soo
CS
Lee
Sang Dae
SD
Hwang
Jae Sam
JS
Kang
Seok Woo
SW
Kim
Hak R
HR
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D018076:DNA, Complementary; D004591:Electrophoresis, Polyacrylamide Gel; D006458:Hemolymph / Q000737:chemistry*; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification; D008297:Male; D008969:Molecular Sequence Data; D009036:Moths; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D019476:Insect Proteins; D011994:Recombinant Proteins