Brett P M, Le Bourdelles B, See C G, Whiting P J, Attwood J, Woodward K, Robertson M M, Kalsi G, Povey S, Gurling H M
University College London Medical School, Academic Department of Psychiatry.
Ann Hum Genet. 1994 May;58(2):95-100. doi: 10.1111/j.1469-1809.1994.tb01879.x.
A cDNA clone of the NMDAR1 (isoform E) has been used to screen both lambda and cosmid genomic libraries. A genomic phage clone was identified and sequenced and was found to contain some of the 3' coding regions of the GRIN1 gene. This clone was used to localize the gene using fluorescent in situ hybridization (FISH) to normal chromosomes, and also to a lymphoblastoid cell line containing a translocation involving chromosomes 9 and 15. FISH localized the gene to chromosome 9q34.3. The clone was used to screen a panel of genomic DNAs cut with 20 restriction enzymes. A VNTR sequence 5' to the gene, which was polymorphic for a number of restriction enzymes, was detected. A PvuII fragment of the genomic clone was found to detect the VNTR on Southern hybridization. The polymorphic VNTR marker was mapped against chromosome 9q34 markers using linkage analysis in the CEPH families. The GRIN1 gene was linked to D9S7 with a maximum lod score of 20.09 at zero recombination fraction in males and 0.03% recombination in females.
利用NMDAR1（异构体E）的一个cDNA克隆筛选λ噬菌体基因组文库和黏粒基因组文库。鉴定并测序了一个基因组噬菌体克隆，发现其包含GRIN1基因的一些3'编码区。该克隆用于通过荧光原位杂交（FISH）将基因定位到正常染色体以及一个包含涉及9号和15号染色体易位的淋巴母细胞系上。FISH将该基因定位到9号染色体q34.3区域。该克隆用于筛选一组用20种限制性酶切割的基因组DNA。在该基因5'端检测到一个VNTR序列，它对多种限制性酶具有多态性。在Southern杂交中发现基因组克隆的一个PvuII片段可检测到该VNTR。利用CEPH家系中的连锁分析，将多态性VNTR标记与9号染色体q34标记进行定位。在男性零重组率时，GRIN1基因与D9S7连锁，最大优势对数得分为20.09；在女性中重组率为0.03%。
