Mycobacterium avium subspecies paratuberculosis (MAP) is the causative pathogen for paratuberculosis, which is a chronic inflammation of the small intestine in ruminants and some wild animals. It affects negatively on the economics of dairy operations worldwide and has a zoonotic concern for its potential relationship to Crohn's disease in humans. In this study, we used different approaches to investigate genetic markers associated with MAP infection on bovine chromosome 7. The interleukin-2-inducible T-cell kinase (ITK) gene is a non-receptor tyrosine kinase expressed in T cells, which also is involved in regulation and development of immune cells and some inflammatory diseases. The gene was suggested in a previous GWAS analysis as a positional candidate gene located on BTA7 for MAP infection. In this study, bovine ITK was sequenced and seventeen identified SNPs were genotyped in 1419 Holstein and Jersey cows. Association analysis revealed no significantly associated SNP in the bovine ITK gene using either single marker or haplotype analyses. In a complementary analysis, Holstein genotypes were imputed from 50 K SNPs to the full genome sequence of BTA7. Single SNP association tests identified two SNPs at 15 Mb (p = 5 × 10-7) significantly associated with MAP infection. Our results suggest an additional region of BTA7 contributes to susceptibility to MAP infection in cattle, relative to our previous report, and further investigations are required.