The invention provides an Escherichia coli bi-component regulation and control system membrane protein ZraS mutant, a gene for coding the mutant, a recombinant vector, a preparation method and an application of the Escherichia coli bi-component regulation and control system membrane protein ZraS mutant. The mutant comprises one or more of S154T, A214S, Q313R and T415A, and the mutant comprises one or more of S154T, A214S, Q313R and T415A. The preparation method of the mutants comprises the following steps: constructing the SP-ZraS bacteriophage; alternately evolving the substrate specificity of the SP-ZraS bacteriophage on the lead ions through positive screening and negative screening by adopting a bacteriophage-assisted continuous directed evolution (PACE) method, so as to obtain an evolutionary sample; performing amino acid sequencing analysis on the evolutionary sample to obtain mutation site information; and constructing an expression vector of the membrane protein ZraS mutant according to mutation site information, recording the expression vector as pZraS, and performing expression to obtain the membrane protein ZraS mutant. The response sensitivity of the mutant to metal lead ions is remarkably improved compared with that of a wild type system, the regulation and control range to lead is enlarged, and the detection limit is as low as 1 mu M; meanwhile, when the ion concentration range is 0-10 mu M, the specific and sensitive detection on the heavy metal lead ions can be realized.