Abstract: A tobacco arsenic transport gene NtNIP7-1 and a cloning method and application thereof are disclosed. A nucleotide sequence of the tobacco arsenic transport gene NtNIP7-1 is shown as SEQ ID: No. 1, and an encoded amino acid sequence thereof is shown as SEQ ID: No. 2. The cloning method of the tobacco arsenic transport gene NtNIP7-1 includes (S1) extracting RNA in tobacco, performing reverse transcription, and obtaining a first-strand cDNA; and (S2) taking the first-strand cDNA obtained by the reverse transcription as a template, synthesizing a specific primer according to sequences of the NtNIP7-1 gene, performing PCR amplification, recovering and purifying a product of the PCR amplification, and sequencing. In the present invention, inhibition of the expression of the tobacco endogenous gene NtNIP7-1 in tobacco plants is able to significantly reduce the arsenic content of tobacco leaves, and has broad application prospects in the field of low arsenic content breeding.