|Van Santen, Vicky|
Submitted to: Genbank
Publication Type: Other
Publication Acceptance Date: 9/14/2008
Publication Date: 9/22/2008
Citation: Xu, D., Panangala, V.S., Van Santen, V.L., Dybvig, K., Abernathy, J.W., Klesius, P.H., Russo, R., Liu, Z. 2008. An immobilization antigen gene of the fish-parasitic protozoan Ichthyophthirius multifiliis strain ARS-6. Genbank. Accession Nos. FJ012354, FJ19440.
Technical Abstract: Ichthyophthirius multifiliis (Ich) is a severe fish parasite that causes ‘white spot’ disease in many freshwater fish and leads to high mortality. The antigens on the parasite surface are involved in the antibody-mediated immobilization and hence designated as immobilization antigens (i-antigens). The nucleotide sequence of an i-antigen gene of Ichthyophthirius multifiliis strain ARS-6 was deduced. The i-antigen coding sequences and flanking sequences were determined from three overlapping PCR products. The amino terminal sequences of purified i-antigen peptide of strain ARS-6 was deduced by mass spectrometry and a partial sequence comprised of a 1.1 kb (central region) coding sequence was determined by PCR amplification of ARS-6 genomic DNA using primers based on previous i-antigen gene sequence in the GenBank. The 5’ portion of the coding sequence and 269 nucleotides of upstream sequence were determined from a PCR product generated by inverse PCR using primers based on the sequence determined from the central portion. The 3’ portion of the coding sequence plus 49 nucleotides of downstream sequences through the polyadenylation site were determined from an amplified and cloned cDNA fragment generated by a rapid amplification of cDNA ends (RACE) procedure. Two overlapping nucleotide sequences of 1627-bp and 1383-bp were obtained, trimmed and aligned to obtain a complete i-antigen gene sequence of ARS-6.