Table 1.

Parallel Transfer of the Chloramphenicol Acetyl Transferase (CAT) Gene into 12 Destination Vectors

Function of Destination Vector1	Colonies2	Control3	Correct4
Expression of native protein in E. coli	15,000	0	6/6
His6-fusion protein in E. coli	10,650	0	6/6
GST-fusion protein in E. coli	9200	0	6/6
Thioredoxin-fusion protein in E. coli	11,000	0	6/6
Sequencing, probe synthesis, (+) strand	13,950	0	6/6
Sequencing, probe synthesis, (−) strand	8950	0	6/6
Transient expression of native protein in mammalian cells	7950	0	6/6
Stable expression of native protein in mammalian cells	10,500	1100	6/6
Expression of native protein in insect cells	7800	15	6/6
Native protein in mammalian cells. Semliki Forest Virus vector	4150	0	6/6
His6-fusion protein in insect cells	6350	30	6/6
Tetracycline-regulated expression of native protein in mammalian cells	11,650	0	6/6

¹Standard vectors were converted to Destination Vectors by inserting a cassette containing the motifs attR1–Cm^R–ccdB–attR2. 

²Aliquots (20 or 200 μL) of the 1 ml expression mixture were plated on ampicillin. Colonies are expressed as the number calculated for mL. Cells were competent at 1 × 10⁸/μg pUC. 

³Identical reactions except the CAT Entry Clone DNA was omitted. 

⁴Based on miniprep DNAs (four colonies) and restriction digests (two colonies) from random colonies.