The invention relates to a multicistron 
plasmid type reverse transcription 
virus vector and a construction method thereof, and belongs to the field of 
gene engineering. The multicistron 
plasmid type reverse transcription 
virus vector is obtained through a 
gene recombination method, and the method comprises the steps that a 
recombinase gene, a simian 
virus large T 
antigen gene, a 
human telomerase gene which are fused with an 
estrogen receptor and five cistrons of a strengthened 
green fluorescent protein gene fused with 
thymidine kinase are inserted into the reverse transcription virus vector, and the a recombinant 
plasmid is obtained. Compared with the prior art, by means of an immortalized 
cell line established through the plasmid type reverse transcription virus vector, non-transformed normal cells can be obtained after 
tamoxifen drug treatment, and meanwhile by means of a 
thymidine kinase suicide gene mechanism, non-returned cells can be made to suicide after 
ganciclovir drug treatment; therefore, complete non-transformed normal cells are obtained. The multicistron plasmid type reverse transcription virus vector and the construction method thereof have the advantages that the 
biological safety of the recombinant reverse transcription virus vector is guaranteed, high expression of multicistron selectivity is promoted, and follow-up screening of a positive 
cell group and detection of multicistron expression are facilitated.