抗体多样性是如何产生的.pptVIP

2021/9/12 * The one-turn/two-turn rule (12/23 rule) - recombination occurs only between a segment with a 12 bp spacer and a segment with a 23 bp spacer. Read Kuby page 113: Recombination Signal Sequences Direct Recombination 2021/9/12 * Kuby Figure 5-6 2021/9/12 * Recombination Activating Genes (encode RAG-1 and RAG-2) RAG-1 and RAG-2 mediate recognition of signal sequences and rearrangement of DNA segments. Mice deficient in RAG-1 or RAG-2 are unable to rearrange heavy or light chain genes. These mice have no mature B cells. -- B cells will not mature if they cannot express a BCR. 2021/9/12 * RAG-deficient mice also lack mature T cells. -- The T cell receptor is also encoded by loci containing gene segments that must be rearranged before the TCR can be expressed. T cells will not mature if they cannot express a TCR. SCID mice also have a defect that affects rearrangement of BCR and TCR loci. They also have no mature T or B cells. 2021/9/12 * Flow cytometry of normal vs. RAG-1 deficient mice: Lymph node cells FITC anti-CD19 (B cell marker) and PE anti-CD3 ( T cell marker) Normal mice RAG-1 - deficient mice 2021/9/12 * 2021/9/12 * 5 3 3 5 5 3 2021/9/12 * 2021/9/12 * A G C T T A T A 2021/9/12 * Terminal deoxynucleotidyl transferase (Tdt) An enzyme that randomly adds in nucleotides during joining of heavy chain (NOT light chain) segments. 2021/9/12 * Read Kuby pages 113-115: Gene Segments Are Joined by Recombinases 2021/9/12 * Productive and nonproductive rearrangements Joining of segments is not precise and may result in loss of the correct reading frame. This may lead to introduction of stop codons -- nonproductive rearrangements. Read Kuby page 115: Ig-Gene Rearrangements May Be Productive or Nonproductive Kuby Figure 5-9 2021/9/12 * Two alleles are available at each locus (maternal and paternal). A B cell expresses only one heavy and light chain allele. - ALLELIC EXCLUSION 2021/9/12 * First, one allele of the heavy chain is rearranged. If