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. 2018 Aug 2;42(4):1977–1986. doi: 10.3892/ijmm.2018.3803

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Copyright: © Fang et al.

This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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Successful construction of the pSEB-CD plasmid. (A) Construction process and structure of the pSEB-CD plasmid. A ~1,300 bp PCR product of full-length CD gene was digested with BamHI/SalI and then directionally cloned into the BglII/SalI-digested pSEB-C3F retroviral vector to construct pSEB-CD plasmid. (B) Positive recombinants were selected by colony PCR, colonies with positive specific PCR products were amplified for plasmid extraction. Lane M, DNA marker; lane 2, 4, 10, 11, 13, 16, 17, 19, positive PCR products specific for CD genes. (C) The constructed pSEB-CD plasmids were confirmed by full-length PCR and enzyme digestion. Lane M1, λ-HindIII marker; lanes 1 and 2, pSEB-CD plasmid was digested with SalI and EcoRI; lanes 3 and 4, full-length CD gene was amplified from pSEB-CD plasmid; lane M2, D2000 marker. (D) The target gene was confirmed to be successfully cloned in the pSEB-C3F vector using gene sequencing. CD, cytosine deaminase; PCR, polymerase chain reaction.