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HLA-JY328：一个多态性HLA I类基因的定位研究与表达

HLA-JY328: mapping studies and expression of a polymorphic HLA class I gene.

作者信息

Duceman B W, Ness D, Rende R, Chorney M J, Srivastava R, Greenspan D S, Pan J, Weissman S M, Grumet F C

出版信息

Immunogenetics. 1986;23(2):90-9. doi: 10.1007/BF00377967.

DOI:10.1007/BF00377967
PMID:3007345
Abstract

The JY328 clone was identified in a human genomic library using cDNA corresponding to mRNA for HLA-B7 as a probe. The L/328 cell line was established by cotransformation of mouse Ltk- cells with the herpes thymidine kinase gene and clone JY328. On Northern blots, RNA from L/328 strongly hybridized to an HLA class I probe, and an antigen was recognized by an anti-HLA class I framework antibody on the cell surface. A DNA probe corresponding to a segment of intron 7 was developed by comparing the nucleotide sequence of clone JY328 with that of other HLA class I-type genes. Using the radiolabeled probe to screen Southern blots of DNA from families with siblings exhibiting intra-HLA recombinations, a restriction fragment length polymorphism was revealed--a 1.4 kb BstE II band not present in all individuals. A corresponding fragment was apparent in the base sequence of clone JY328. The occurrence of this band on Southern blots established that JY328 maps distinct from and centromeric to the HLA-C locus and near to the HLA-B locus. Antibody absorption studies and cytotoxicity tests indicated that the JY328 gene product was not an HLA-B antigen but that it did specifically absorb CW7-specific antibody. In sum, these results suggest a novel, polymorphic HLA class I gene which expresses a product serologically similar to HLA-Cw7 but which does not map within the corresponding locus.

摘要

使用与HLA - B7的mRNA对应的cDNA作为探针，在人类基因组文库中鉴定出JY328克隆。通过用疱疹胸苷激酶基因和克隆JY328共转化小鼠Ltk - 细胞，建立了L/328细胞系。在Northern印迹上，来自L/328的RNA与HLA I类探针强烈杂交，并且细胞表面上的一种抗原被抗HLA I类构架抗体识别。通过比较克隆JY328与其他HLA I类基因的核苷酸序列，开发了一种与内含子7的一段相对应的DNA探针。使用放射性标记的探针筛选来自具有HLA内重组的同胞家庭的DNA的Southern印迹，揭示了一种限制性片段长度多态性——并非所有个体中都存在的1.4 kb BstE II条带。在克隆JY328的碱基序列中可以看到一个相应的片段。Southern印迹上该条带的出现确定了JY328定位于与HLA - C基因座不同且在着丝粒方向，并且靠近HLA - B基因座。抗体吸收研究和细胞毒性试验表明，JY328基因产物不是HLA - B抗原，但它确实特异性吸收CW7特异性抗体。总之，这些结果提示了一种新的多态性HLA I类基因，其表达的产物在血清学上与HLA - Cw7相似，但不在相应基因座内定位。

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引用本文的文献

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J Exp Med. 1995 Oct 1;182(4):1005-18. doi: 10.1084/jem.182.4.1005.
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HLA-C genes are transcribed in HLA-C blank individuals.

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