Fig. 4.
Fig. 4. Construction of the expression vector with ALAD cDNA. / Recombinant 1: Normal ALAD cDNA cloned from EBV-transformed lymphoblastoid cells of the patient (allele 2). Recombinant 2: Normal ALAD cDNA introduced in the opposite direction into the pdKCR-Neo vector. Recombinant 3: G177 to C transition ALAD cDNA, reconstituted by replacing EcoRI-NcoI fragment (637 bp) from normal ALAD cDNA with theEcoRI-NcoI fragment (444 bp) of G177to C/G397 to A transition of ALAD cDNA. Recombinant 4: G397 to A transition of ALAD cDNA, reconstituted by replacing EcoRI-NcoI fragment (444 bp) from normal ALAD cDNA with the EcoRI-NcoI fragment (637 bp) of G177 to C/G397 to A transition of ALAD cDNA. Recombinant 5: G177 to C and G397 to A transition of ALAD cDNA, cloned from lymphoblastoid cells of the proband (allele 1). These recombinant cDNA were introduced into theEcoRI site of the third exon of rabbit β-globin in the pdKCR-Neo expression vector.

Construction of the expression vector with ALAD cDNA.

Recombinant 1: Normal ALAD cDNA cloned from EBV-transformed lymphoblastoid cells of the patient (allele 2). Recombinant 2: Normal ALAD cDNA introduced in the opposite direction into the pdKCR-Neo vector. Recombinant 3: G177 to C transition ALAD cDNA, reconstituted by replacing EcoRI-NcoI fragment (637 bp) from normal ALAD cDNA with theEcoRI-NcoI fragment (444 bp) of G177to C/G397 to A transition of ALAD cDNA. Recombinant 4: G397 to A transition of ALAD cDNA, reconstituted by replacing EcoRI-NcoI fragment (444 bp) from normal ALAD cDNA with the EcoRI-NcoI fragment (637 bp) of G177 to C/G397 to A transition of ALAD cDNA. Recombinant 5: G177 to C and G397 to A transition of ALAD cDNA, cloned from lymphoblastoid cells of the proband (allele 1). These recombinant cDNA were introduced into theEcoRI site of the third exon of rabbit β-globin in the pdKCR-Neo expression vector.

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