Objective：To constract the Rv2647-deleted strain and Rv2647-complemented strain of Mycobacterium tuberculosis（M. tb） and Rv2647-overexpressing Mycobacterium smegmatis（Ms：：Rv2647）by phage recombination technique，and to evaluate the effect of M. tb Rv2647 protein on lung injury in model mice. Methods：A homologous exchange site was constructed and integrated into the phage genomes of M. tb，producing phagemids that were introduced into Mycobacterium smegmatis（Ms）to create a recombinant phage with a homologous exchange site. High-titer recombinant phages were amplified in vitro and transfected into M. tb（H37Rv），followed by static culture at 37 ℃ for 28 d. The single clone was selected and verified by PCR，yeilding the Rv2647 - deleted strain （H37RvΔRv2647）. The Rv2647 gene was amplified by PCR and was integrated into the multiple clone sites of vector pMV361 and pMV261 through seamless cloning to obtain the positive plasmids，which were transfected into H37RvΔRv2647 and Ms to obtain the Rv2647 - complemented strain（H37RvΔRv2647：：Rv2647）and Rv2647 - overexpressing Ms（Ms：：Rv2647），respectively. The suspension of H37Rv，H37RvΔRv2647，H37RvΔRv2647：：Rv2647，Ms，and Ms：：Rv2647 were used to infect C57BL/6 mice via tracheal injection. The survival rates，lung tissue bacterial load，and pathological damage of lung tissue in model mice were compared at 7 d and 30 d，respectively. Results：The PCR showed that Rv2647 gene was missing in the H37RvΔRv2647，while it was present in the H37RvΔRv2647：：Rv2647 and Ms：：Rv2647. The 30-day survival rates of model mice infected with H37RvΔRv2647，H37Rv，and H37RvΔRv2647：：Rv2647 were 100.00%，83.33%，and 83.33%，respectively；The 7-day survival rates of model mice infected with Ms and Ms：：Rv2647 were 100.00% and 83.33%，respectively. The lung bacterial load（lgCFU）of model mice in the H37RvΔRv2647 group（3.40±0.18）was significantly lower than that of the H37Rv group（3.86±0.15），P ＜ 0.001）and the H37RvΔRv2647：：Rv2647 group（3.80±0.16），P ＜ 0.01）；The inflammation area（%）in lung tissues of the H37RvΔRv2647 group（4.37±3.06）was significantly lower than that of the H37Rv group（62.76±14.24），P ＜ 0.001）and the H37RvΔRv2647：：Rv2647 group（67.37±0.45），P ＜ 0.001）. The lung bacterial load（lgCFU）of the Ms group（2.53±0.16）was significantly lower than that of the Ms：：Rv2647 group（2.81±0.13）， P ＜ 0.01）；The inflammation area（%）in lung tissue of the Ms group（5.71±1.29）was significantly lower than that of the Ms：：Rv2647 group（33.13 ± 13.84），P ＜ 0.05）. Conclusion：The Rv2647 - deleted strain（H37RvΔRv2647）and Rv2647 - complementary strain （H37RvΔRv2647：：Rv2647）of M. tb and Rv2647-overexpressing Ms（Ms：：Rv2647）were successfully constructed. Rv2647 protein may aggravate lung injury via inhibiting host clearance of M. tb.