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In this study, pig ST6GalNAc IV gene was cloned from pig liver tissue and characterized. The pST6GalNAc IV cDNA has the open reading frame (ORF) containing 909 bp encoded 302 amino acids. Entire ORF of pST6GalNAc IV containing sialylmotif, ‘L’- (Large), ‘S’- (Small) and ‘VS’- (Very small) motif has a higher degree of sequence homology with Homo sapiens (90%), Pan troglodytes (91%) and Mus musculus (87%). Expression of pST6GalNAc IV mRNA in various pig tissues was identified by Reverse transcription polymerase chain reaction (RT-PCR) analysis. The pST6GalNAc IV mRNA was highly expressed in tongue, muscle and heart, but not expressed in pancreas.We also established pST6GalNAc IV-transfected PK15 stable cells for observing functional characterization of pST6GalNAc IV gene. Glycosylation pattern of pST6GalNAc IV-transfected PK15 cells was detected by flow cytometry and Immunofluorescence analysis with peanut agglutinin lectin (PNA) and maackia amurensis lectin II (MAL II). These results revealed that Neu5Ac-α2,3Gal-1,3(Neu5Ac α 2-6)GalNAc tetrasaccharide structures found on O-glycan were newly produced by pST6GalNAc IV, whereas Gal-1,3GalNAc disaccharide structures found on O-glycan were decreased by pST6GalNAc IV.