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. 2001 Feb;123(2):340–345. doi: 10.1046/j.1365-2249.2001.01437.x

Fig. 1.

Fig. 1

Cloning and expression of the TCR scFv. TCR α genes of Vα12+ T cells and TCR β genes of the two clonally expanded T cells (A22 and P39) were cloned for single strand conformation polymorphism (SSCP), nucleotide sequencing and/or expressing the TCR scFv protein. (a) Location of each primer is indicated by arrows. SS, Signal sequence. (b) The TCR genes of A22 were subcloned to plasmid vectors for expressing the scFv protein. The TCR α and β genes were subcloned to VL and VH sites of 9F12-encoding pCANTAB5E, respectively. The entire gene of TCR α/linker/TCRβ/E-Tag was transferred to EcoRI/BamHI-digested pMAL-c vector. The TCR scFv was expressed as a fusion protein with maltose binding protein (MBP) and E-Tag. After the MBP was cleaved by Factor Xa, the TCR scFv/E-Tag was purified on an affinity column using the anti-E-Tag antibody.