Table 1. Efficiency of the mini-F vector transposition in E. coli and the vector self-excision upon virus reconstitution.
| Efficiency of the mini-F transposition and the mini-F self-excision | ||||||
| target | bacteria positive/tested | virus passage | viruses positive/tested | |||
| mini-F excisiona | mini-F transpositionb | mini-F detectiond | mini-F excisionf | |||
| Colony-PCR | RFLP | sequencing | PCR (experiments) | PCR | sequencing | |
| ORF22 | 3/23 | 2/3 | 2/2c | 4 (6) | 6/6 | 2/2 |
| ORF50 | 15/23 | 8/8 | 6/6c | 4 (4) | 4/4 | 2/2 |
| ORF54 | 15/23 | 8/8 | 6/6c | 4 (1) | 1/1 | 1/1 |
| ORF62 | 5/96 | 2/5 | 2/2 | 4; >7e (2) | 1/2e | 1/1 |
| ORF71 | 8/96 | 2/8 | 2/2 | 5; 6 (2) | 2/2 | 2/2 |
| Junction | 9/168 | 3/9 | 1/1 | 2 (8) | 8/8 | 16/16 |
a
Colony-PCR screening for determination of the mini-F excision from the US unique AvrII site.
b
Confirmation of the mini-F transposition into the given target site by RFLP and PCR or sequencing analysis.
c
Only the repair of the unique AvrII site was investigated via sequencing.
d
Average number of the last virus passage in which the mini-F sequences were detected by PCR after virus reconstitution. Numbers of the performed independent transfection experiments are given in brackets.
e
Detection of mini-F until passage 7 and beyond.
f
Confirmation of the mini-F excision in derived virus progenies by PCR or sequencing.