Frequency and range of mutations in a 500-bp-long region in the intron downstream of the rearranged VHDHJH4 joints of GC B cells derived from pairs of M17+/+ and M17–/– mice
Organ and genotype . | No. clones . | No. mutated clones . | Range, mutations/clone . | No. mutations/no. bp (%) . |
---|---|---|---|---|
Spleen | ||||
M17+/+ | 35 | 25 | 1-14 | 102/12 248 (0.83) |
M17-/- | 56 | 31 | 1-12 | 117/15 500 (0.75) |
Peyer patches | ||||
M17+/+ | 26 | 25 | 1-20 | 196/12 500 (1.57) |
M17-/- | 30 | 25 | 1-25 | 213/12 447 (1.71) |
Organ and genotype . | No. clones . | No. mutated clones . | Range, mutations/clone . | No. mutations/no. bp (%) . |
---|---|---|---|---|
Spleen | ||||
M17+/+ | 35 | 25 | 1-14 | 102/12 248 (0.83) |
M17-/- | 56 | 31 | 1-12 | 117/15 500 (0.75) |
Peyer patches | ||||
M17+/+ | 26 | 25 | 1-20 | 196/12 500 (1.57) |
M17-/- | 30 | 25 | 1-25 | 213/12 447 (1.71) |
Naive M17-/- B cells had a 0.03% mutation frequency.
GC B cells were isolated 14 days after immunization with NP-CG. Following cell lysis, a 600-bp fragment was PCR amplified from 40 000 cell equivalents using a primer pair that anneals in the framework 3 region of most J558 V genes and in the intron downstream of JH4 gene segment.