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題名 | Human α-L-Iduronidase (IDUA) Gene: Correlation of Polymorphic DNA Haplotype and IDUA Activity in Chinese Population=人類α-L-Iduronidase (IDUA)基因:中國人族群的多型性DNA單套型與IDUA活性的相關性 |
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作者 | 李桂楨; 王政光; 黃淑芬; 戴坤蓉; Lee-chen, Guey-jen; Wang, Cheng-kuang; Huang, Shu-fen; Day, Kuen-rong; |
期刊 | Proceedings of the National Science Council : Part B, Life Science |
出版日期 | 19980100 |
卷期 | 22:1 1998.01[民87.01] |
頁次 | 頁31-38 |
分類號 | 361.1925 |
語文 | eng |
關鍵詞 | IDUA酵素活性; 中國人族群; 多型性DNA單套型; IDUA polymorphism; Haplotype; α-L-iduronidase activity; |
中文摘要 | 本論文在探討中國人族族的多型性 DNA 單套型與 IDUA 酵素活性的相關性。我們 自正常人族群逢機取樣 85 年,萃取其基因組 DNA, 放大包含 A8、Q33H (表現子 1 )、 R105Q (表現子 3 )、 A361T (表現子 8 )或 V454I (表現子 9 )等多型性點的 DNA 片段, 再以限制片段長度多型性( RFLP )或對偶基因專一的寡核甘酸引子( ASO ),來 檢測各樣品的多型性基因型。我們並自上述各血液樣品中分離白血球,測其 IDUA 酵素活性 。 結果發現各樣品的 IDUA 酵素活性差異頗大(約介於 50~300 nmol/h/mg cell protein 間),其平均值約 156 nmol/h/mg cell protein。在酵素活性高於 110 %平均值的樣品群 中, Q33H 的不具多型性的對偶基因 "1" 及 R105Q、A361T、V454I 的多型性對偶基因 "2" 的頻率較酵素活性低於 90 %平均值的樣品群為高。 單套型的連鎖不平衡分析顯示,R105Q 、 A361T 和 V454I 的多型性對偶基因間呈現連鎖關係。 當檢測 Q33H、R105Q、 A361T 及 V454I 的單套型時,單套型 1,2,2,2 ( Gln33, Gln105, Thr361, Ile454 )和 IDUA 酵素 活性有相關性。帶有 R105Q、A361T 或 V454I 多型性的 IDUA cDNA,於 COS-7 細胞中所表 現的酵素活性, 均較正常 cDNA 高。若將 Gln105、Thr361、Ile454 等和酵素活性正相關的 多型性集中於一 cDNA 上,則或可表現出一具高活性的 IDUA 蛋白,應用於 MPSI 患者的酵 素替換治療上。 |
英文摘要 | The correlation of polymorphic DNA haplotype of the α -L-iduronidase (IDUA) gene and IDUA activity in Chinese subjects was investigated. Genomic DNA extracted from 85 randomly sampled normal individuals was used to amplify fragments containing the polymorphic change site A8, Q33H (exon 1), R105Q (exon 3), A361T (exon 8), or V454I (exon 9). The PCR amplified products were analyzed by means of restriction fragment length polymorphism (REFLP) or allede specific oligonucleotide (ASO) hybridization. Leukocytes were isolated from the above 85 samples, and their IDUA activities were determined. A wide range of IDUA activity (50~300 nmol/h/mg cell protein) with an average of 156 nmol/h/mg cell protein was observed. When the allele frequency was compared between individuals with IDUA activity below 90% of above 110% of the average, a bias toward the common allele "1" of Q33H (Gln33) was detected in individuals with higher IDUA activity. Conversely, the polymorphic allele "2" of R105Q (Gln105), A361T (Thr361), and V454I (Ile454) was found in the higher IDUA activity group. Linkage disequilibrium analysis of the haplotype data revealed strong nonrandom association among the polymorphic alleles of R105Q, A361T, and V454I. Of the haplotypes constructed by Q33H, R105Q, A361T, and V454I, a positive correlation between haplotype 1,2,2,2 (Gln33, Gln105, Thr361, Ile454) and IDUA activity was observed. The IDUA activity was found to increase with Gln105, Thr361, or Ile454 polymorphic changes by mutagenesis and expression of IDUA cDNA in COS-7 cells. By combining the positive effect of Gln105, Thr361, and Ile454 in one cDNA construct, it may be possible to produce a high avtivity IDUA protein for MPSI enzyme replacement therapy. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。