Cloning, nucleotide sequence and phylogenetic analyses, and tissue-specific expression of the transferrin gene in Cirrhinus mrigala infected with Aeromonas hydrophila

Abstract

Transferrin partial complementary DNAs were cloned from the livers of five species in four genera of Indian carps (Indian major carp species: Labeo rohita, Catla catla and Cirrhinus mrigala; medium carp: Puntius sarana; minor carp: Labeo bata) subsequent to polymerase chain reaction amplification with published heterologous primers or self-designed primers derived from conserved regions of transferrin cDNA sequences. The partial transferrin cDNAs of the five species of carps had sizes from 624 to 633bp (487bp for L. rohita) and encoded an open reading frame consisting of 206-211 (162 for L. rohita) amino acids. The alignments of carp cDNA sequences showed 85-97% homology and 71-93% homology in deduced amino acid sequences. A phylogenetic tree of amino acid sequences of transferrin cDNAs from carps showed that the relationship among the four genera of Indian carps is well correlated with that derived from classic morphologic analyses. The hypothesized cleavage site and interdomain bridge of transferrin molecule were predicted for the above carp species and interestingly the cleavage site amino acid sequence was found to be conserved among all the carps. To study the tissue-specific expression of the transferrin gene, various tissues (liver, kidney, spleen, brain, muscle, testis, heart, intestine, gill and fin) from apparently healthy (control), moribund and survived C. mrigala experimentally infected with Aeromonas hydrophila infection were analyzed. Transferrin mRNA was detected only in liver RNA and to lesser extent in brain tissue out of the 10 tissues analyzed irrespective of bacterial infection.