In this study, the relative expression of NtMTP1 was determined by quantitative realtime PCR (qRTPCR). The results showed that NtMTP1 gene was highly expressed in flowers and leaves. After treatment of 400 μmol/L ZnSO₄ for 48 h, the expression of NtMTP1 increased to the maximum, which was 3.81 folds of the control. Plantexpression vector pBI12135S∶∶MTP1 was constructed, and the recombinant plasmid was introduced into Nicotiana tabacum L. cv NC89 by means of Agrobacteriumtumefaciens mediated transformation. The qRTPCR revealed a distinct increase of endogenous NtMTP1 expression level in the overexpressing transgenic lines, which were 10.42, 7.61 and 11.84 folds of that of wild plants, respectively. Seeds of wild type and transgenic plants were treated with different concentrations of ZnSO₄. The results showed that overexpression of NtMTP1 conferred enhanced zinc tolerance. These results provide an important basis to illuminate the function of NtMTP1 gene in Zn²⁺ transport in tobacco.