Paramyosin from the parasitic mite Sarcoptes scabiei: cDNA cloning and heterologous expression.
The burrowing mite Sarcoptes scabiei is the causative agent of the highly contagious disease sarcoptic mange or scabies. So far, there is no in vitro propagation system for S. scabiei available, and mites used for various purposes must be isolated from infected hosts. Lack of parasite-derived material has limited the possibilities to study several aspects of scabies, including pathogenesis and immunity. It has also hampered the development of high performance serological assays. We have now constructed an S. scabiei cDNA expression library with mRNA purified from mites isolated from red foxes. Immunoscreening of the library enabled us to clone a full-length cDNA coding for a 102.5 kDa protein. Sequence similarity searches identified the protein as a paramyosin. Recombinant S. scabiei paramyosin expressed in Escherichia coli was recognized by sera from dogs and swine infected with S. scabiei. We also designed a small paramyosin construct of about 17 kDa that included the N-terminal part, an evolutionary variable part of the helical core, and the C-terminal part of the molecule. The miniaturized protein was efficiently expressed in E. coli and was recognized by sera from immunized rabbits. These data demonstrate that the cDNA library can assist in the isolation of important S. scabiei antigens and that recombinant proteins can be useful for the study of scabies.
Mattsson
J G
JG
Department of Parasitology (SWEPAR), National Veterinary Institute, Uppsala, Sweden. Jens.Mattsson@sva.se
Ljunggren
E L
EL
Bergström
K
K
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D004926:Escherichia coli; D005260:Female; D005589:Foxes; D005786:Gene Expression Regulation; D015723:Gene Library; D008297:Male; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000737:chemistry; D012520:Sarcoptes scabiei / Q000235:genetics*; D014335:Tropomyosin / Q000235:genetics*
D018076:DNA, Complementary; D012333:RNA, Messenger; D014335:Tropomyosin