Isolation and identification of an actin gene from Babesia gibsoni.
Actin is a ubiquitous and highly conserved microfilament protein that is hypothesized to play a mechanical force-generating role in the unusual gliding motility of sporozoan zoites and their active penetration of host cells. We have identified and isolated an actin gene from a Babesia gibsoni cDNA library by random sequencing. The complete nucleotide sequence of the actin gene is 1,243 bp; a single open reading frame encodes a polypeptide of 377 amino acid residues. The deduced amino acid sequence showed a high homology with actins from other species, especially with reported apicomplexan protozoans. The antiserum against recombinant actin expressed in Escherichia coli recognizes a 42-kDa native protein, which is consistent with its expected size. Immunofluorescence and confocal microscopic observation revealed that the protein is diffusely distributed throughout the B. gibsoni parasites.
Zhou
Jinlin
J
National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan.
Huang
Bing
B
Suzuki
Hiroshi
H
Fujisaki
Kozo
K
Igarashi
Ikuo
I
Xuan
Xuenan
X
D000199:Actins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis; D001403:Babesia / Q000235:genetics*; D001483:Base Sequence; D015153:Blotting, Western / Q000379:methods; D016054:DNA, Protozoan / Q000737:chemistry; D004285:Dogs; D004926:Escherichia coli / Q000235:genetics; D005455:Fluorescent Antibody Technique / Q000379:methods; D015870:Gene Expression; D015723:Gene Library; D017125:Genes, Protozoan / Q000235:genetics* / Q000502:physiology; D018613:Microscopy, Confocal / Q000379:methods; D008858:Microscopy, Phase-Contrast / Q000379:methods; D008969:Molecular Sequence Data; D011419:Propidium / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid
D000199:Actins; D000953:Antigens, Protozoan; D016054:DNA, Protozoan; D011994:Recombinant Proteins; D011419:Propidium