The Wellcome Trust Sanger Institute, Hinxton, UK.
Genomics. 2010 Feb;95(2):105-10. doi: 10.1016/j.ygeno.2009.10.004. Epub 2009 Nov 10.
Non-obese diabetic (NOD) mice spontaneously develop type 1 diabetes (T1D) due to the progressive loss of insulin-secreting beta-cells by an autoimmune driven process. NOD mice represent a valuable tool for studying the genetics of T1D and for evaluating therapeutic interventions. Here we describe the development and characterization by end-sequencing of bacterial artificial chromosome (BAC) libraries derived from NOD/MrkTac (DIL NOD) and NOD/ShiLtJ (CHORI-29), two commonly used NOD substrains. The DIL NOD library is composed of 196,032 BACs and the CHORI-29 library is composed of 110,976 BACs. The average depth of genome coverage of the DIL NOD library, estimated from mapping the BAC end-sequences to the reference mouse genome sequence, was 7.1-fold across the autosomes and 6.6-fold across the X chromosome. Clones from this library have an average insert size of 150 kb and map to over 95.6% of the reference mouse genome assembly (NCBIm37), covering 98.8% of Ensembl mouse genes. By the same metric, the CHORI-29 library has an average depth over the autosomes of 5.0-fold and 2.8-fold coverage of the X chromosome, the reduced X chromosome coverage being due to the use of a male donor for this library. Clones from this library have an average insert size of 205 kb and map to 93.9% of the reference mouse genome assembly, covering 95.7% of Ensembl genes. We have identified and validated 191,841 single nucleotide polymorphisms (SNPs) for DIL NOD and 114,380 SNPs for CHORI-29. In total we generated 229,736,133 bp of sequence for the DIL NOD and 121,963,211 bp for the CHORI-29. These BAC libraries represent a powerful resource for functional studies, such as gene targeting in NOD embryonic stem (ES) cell lines, and for sequencing and mapping experiments.
非肥胖型糖尿病(NOD)小鼠由于自身免疫驱动的过程导致胰岛素分泌β细胞逐渐丧失,从而自发地发展为 1 型糖尿病(T1D)。NOD 小鼠是研究 T1D 遗传和评估治疗干预的有价值的工具。在这里,我们通过末端测序描述了源自 NOD/MrkTac(DIL NOD)和 NOD/ShiLtJ(CHORI-29)的细菌人工染色体(BAC)文库的开发和表征,这两个常用的 NOD 亚系。DIL NOD 文库由 196032 个 BAC 组成,CHORI-29 文库由 110976 个 BAC 组成。从将 BAC 末端序列映射到参考小鼠基因组序列来估计 DIL NOD 文库的基因组覆盖深度,在常染色体上为 7.1 倍,在 X 染色体上为 6.6 倍。该文库中的克隆平均插入大小为 150kb,映射到参考小鼠基因组组装(NCBIm37)的超过 95.6%,覆盖了 Ensembl 小鼠基因的 98.8%。按照相同的度量标准,CHORI-29 文库在常染色体上的平均深度为 5.0 倍,X 染色体上的覆盖度为 2.8 倍,X 染色体覆盖度降低是因为该文库使用了雄性供体。该文库中的克隆平均插入大小为 205kb,映射到参考小鼠基因组组装的 93.9%,覆盖了 Ensembl 基因的 95.7%。我们已经鉴定和验证了 DIL NOD 的 191841 个单核苷酸多态性(SNP)和 CHORI-29 的 114380 个 SNP。总共为 DIL NOD 生成了 229736133bp 的序列,为 CHORI-29 生成了 121963211bp 的序列。这些 BAC 文库是功能研究的强大资源,例如在 NOD 胚胎干细胞(ES)系中的基因靶向,以及测序和映射实验。