Table III.
Conserved cRSs from multiple VH gene segments are cleaved in pro–B cells from RAG2:GFP, C57BL/6, and μMT mice
| Position (nt) |
cRS site |
VH gene | VH cRS sequence | Mouse strain |
|---|---|---|---|---|
| 57 | I | IGHV1S2*01 | TCCAACTGCAGCAGCCTGGGGC TGAGCTTGT GAAGCCTGGGGC TTCAGTG | RAG2:GFP |
| 122 | II | IGHV1-56*02 | ATATCCTGCAAGGCTCCTGGCT ACACCTTCA CCAGCCACTGGA TGCAGTG | μMT |
| 122 | II | IGHV1S132*01 | CTGTCCTGCAAGACTTCTGGCT ACACCTTCA CCAGCTACTGGA TTCAGTG | μMT |
| 155 | III | IGHV1-14*01 | AGCTATGTTATGCACTGGGTGA AGCAGAAGC CTGGGCAGGGCC TTGAGTG | μMT |
| 155 | III | IGHV1-22*01 | GACTACAACATGCACTGGGTGA AGCAGAGCC ATGGAAAGAGCC TTGAGTG | C57BL/6 |
| 155 | III | IGHV1-39*01 | GGCTACACCATGAACTGGGTGA AGCAGAGCC ATGGAAAGAACC TTGAGTG | C57BL/6 |
| 155 | III | IGHV1-54*01 | AATTACTTGATAGAGTGGGTAA AGCAGAGGC CTGGACAGGGCC TTGAGTG | μMT, C57BL/6 |
| 155 | III | IGHV1-55*01 | AGCTACTGGATAAACTGGGTGA AGCTGAGGC CTGGACAAGGCC TTGAGTG | RAG2:GFP |
| 155 | III | IGHV1-63*01 | AACTACTGGATAGGTTGGGTAA AGCAGAGGT CTGGACATATAC ATGGGTG | RAG2:GFP |
| 155 | III | IGHV1-67*02 | GATTATGCTATGCACTGGGTGA AGCAGAGTC ATGCAAAGAGTC TAGAGTG | μMT |
| 155 | III | IGHV1S1*01 | AGCTACTGGATGCACTGGGTGA AGCAGAGGC CTGGACGAGGCC TCGAGTG | RAG2:GFP |
| 155 | III | IGHV1S2*01 | AGCTACTGGATGCACTGGGTGA AGCAGAGGC CTGGACGAGGCC TTGAGTG | RAG2:GFP |
| 155 | III | IGHV1S10*01 | ACCTACTGGATGAACTGGGTGA AGTAGATGC CTGGACAGGGCC TTGAGTG | μMT, C57BL/6 |
| 155 | III | IGHV1S25*01 | GAGTATATTATACACTGGGTAA AGCAGAGGT CTGGACAGGGTC TTGAGTG | μMT |
| 155 | III | IGHV1S30*01 | AGCTACTACATGCACTGGGTGA AGCAGAGCC ATGGAAAGAGCC TTGAGTG | μMT |
| 155 | III | IGHV1S32*01 | AGCTACTATATACACTGGGTGA AGCAGAGGC CTGGACAGGGAC TTGAGTG | μMT |
| 155 | III | IGHV1S55*01 | AGCTCCTGGATGAACTGGGTGA AGCAGAGGC CTGGACAGGGAC TTGAGTG | C57BL/6 |
| 155 | III | IGHV1S70*01 | AGCTACTGGATAAACTGGGTGA AGCAGAGGC CTGGACAAGGCC TTGAGTG | μMT |
| 155 | III | IGHV1S95*01 | AGCTACTGGATGCACTGGGTGA AGCAGAGGC CTGGACAAGGCC TTGAGTG | μMT, RAG2:GFP |
| 155 | III | IGHV1S130*01 | AGCTCCTGGATGCACTGGGCGA AGCAGAGGC CTGGACAAGGCC TTGAGTG | μMT |
| 181 | III | IGHV1-64*01 | GAGGCCTGGACAAGGCCTTGAG TGGATTGGA ATGATTCATCCT AATAGTG | μMT, C57BL/6 |
| 267 | - | IGHV1S14*01 | AGGGCAAGGCCACAATGACTGT AGACACATC CTCCAGCACAGC CTACGTG | RAG2:GFP |
| 267 | - | IGHV1S130*01 | AGGGCAAGGCCACACTGACTGT AGACACATC CTCCAGCACAGC CTACGTG | μMT |
| 313 | V | IGHV1-4*01 | CATGCAACTGAGCAGCCTGACA TCTGAGGAC TCTGCAGTCTAT TACTGTG | C57BL/6 |
| 313 | V | IGHV1-19*01 | CATGGAGCTCAACAGCCTGACA TCTGAGGAC TCTGCGGTCTAT TACTGTG | μMT |
| 313 | V | IGHV1-48*02 | CATGGAGCTCAGCAGCCTGACA TCTGAGGAC TCTGCAGTCTAT TACTGTG | C57BL/6 |
| 313 | V | IGHV1-60*01 | CATGCAGCTCAGCAGCATGACA TCTGAAGCC TCTGATGACTAT TACTGTG | C57BL/6 |
| 313 | V | IGHV1S126*01 | CATGCAGCTCAGCAGCCTGACA TCTGAGGAC TCTGCGGTCTAT TACTGTG | C57BL/6 |
| 313 | V | IGHV5-6-2*01 | CCTGCAAATGAGCAGTCTGAAG TCTGAGGAC ACAGCCTTGTAT TACTGTG | RAG2:GFP |
The positions and sequences of VH 12-cRS SEs recovered from pro–B cells sorted from the BM of μMT, RAG2:GFP, or C57BL/6 mice are listed. Recovered VH1 and VH5 SE LM-PCR products were cloned into the pCR2.1TOPO vector and sequenced. Ligation of the BW-LC linker directly to the cRS heptamer confirmed RAG1/2-mediated cleavage. The sequences were processed in Vector NTI and analyzed by the IMGT database (http://imgt.cines.fr) and Immunoglobulin BLAST (http://www.ncbi.nlm.nih.gov/igblast) for gene identification. A variety of VH gene segments contain functional cRSs, and most SE products represent site III cRSs. Heptamer and nonamer sequences of VH 12-cRS are in bold.