汉、蒙古族人MBL基因ExonⅠ54位密码子点突变频率及其血浆含量相关性研究  

Correlation between point mutation frequency of MBL coding gene Exon 1 and MBL plasma concentration in Chinese Hans and Mongolia people

作　　者：贾天军[1] 李萍[1] 张庶民[2] 刘洁[1] 赵铁军[1] 张进顺[1] 雷殿良[2] 翟登高[1] 李河民[2] 

机构地区：[1]张家口医学院实验中心,063000 [2]中国药品生物制品检定所血清室,北京100050

出　　处：《中华微生物学和免疫学杂志》2004年第2期115-119,共5页Chinese Journal of Microbiology and Immunology

基　　金：河北省自然科学基金资助 ( 3 0 0 3 77);863博士基金资助 ( 2 0 0 3AA2Z3 5 0 9)

摘　　要：目的　调查健康汉、蒙古族人甘露 (聚 )糖结合凝集素 (MBL)基因 5 4位密码子点突变的情况 ,测定健康汉、蒙古族人血浆MBL含量 ,并对健康汉、蒙古族人MBL基因 5 4位密码子点突变频率与其血浆MBL含量进行相关性分析。方法　PCR扩增 ,测定序列并建立MBL基因点突变检测方法 ,即PCR RFLP方法 (BanⅠ酶切 ) ;用ELISA法测血浆MBL含量。结果　 (1)建立了MBLPCR方法 ,该方法特异性高 ,重复性好 ,最低检出量为 16 0pgDNA。 (2 )MBL的限制性内切酶BanⅠ酶切结果 :通过酶切电泳分析结果显示 ,所扩增标本出现下列 3种情况 :①MBL 5 4位密码子野生型纯合子为 2条带 ,对应的相对分子质量为 2 32bp和 93bp ;② 5 4位密码子突变体纯合子只显示 1条约 32 5bp带 ;③ 5 4位密码子野生型突变体杂合子显示 32 5bp、2 32bp和 93bp 3条带。 (3)通过简单数基因计算MBL基因5 4位密码子点突变的频率 ,健康汉、蒙古族人基因突变频率分别为 0 .2 32 1和 0 .175 7。 (4 )血浆MBL含量的测定 :5 6例健康汉族人血浆MBL含量的平均值为 1998.75 0 μg L ,标准差为 15 0 5 .15 2 ;37例健康蒙古族人血浆MBL含量的平均值为 2 5 2 5 .6 76 μg L ,标准差为 195 5 .188。 (5 )健康汉族人血浆MBL含量与其编码基因ExonⅠ 5 4位密码子的点突变频率 ,Objective To analyze the point mutation of MBL (mannose-binding lectin or mannan-binding lectin) Exon 1 at codon 54 in healthy Chinese Hans and Mongolia people, to measure the plasma levels of MBL, and to analyze the association between gene mutation frequency and the plasma concentrations of MBL in both groups. Methods Blood samples were from collected 56 healthy Hans and 37 healthy Mongolia people; Establishment of PCR amplification and method for the detection of MBL point mutation (PCR-RFLP); Assay for plasma MBL: MBL plasma concentrations were measured using MBL ELISA. Results (1) No bands were found in non-specificity samples (HAV, HBV, HCV and TB) , and the sequences of PCR products were the same as the expected ones by sequencing analysis. (2) Frequency of point mutation in healthy Hans and Mongolia people were 0.2321 and 0.1757 respectively. (3) The average plasma MBL concentration was 1998.750μg/L, standard deviation was 1505.152 in 57 healthy Hans; that of 37 Mongolia people was 2525.676μg/L, standard deviation was 1955.188. (4) Analysis of correlation between MBL concentrations and gene mutation frequency in healthy Hans: Frequency of point mutation was 1.00 when the MBL concentrations were below 100μg/L; frequency of point mutation was 0.4524 when in the range of 100μg /L to 1000μg /L; frequency of point mutation was 0.0156 when over 1000μg /L. (5) Frequency of point mutation was 1.00 when the MBL concentrations were below 100μg /L; frequency of point mutation was 0.4583 from 100μg /L to 1000μg/L; no point mutation was found in over 1000μg/L. Conclusion (1) PCR-RFLP, detecting the point mutation at codon 54 of MBL coding gene, had a high specificity, reproducibility and good sensitivity. (2) The frequency of mutation at codon 54 of MBL coding gene had been determined in both healthy Hans and Mongolia people, the frequency was higher in healthy Hans than in Mongolia people, but no statistical significant(χ 2=0.8574,P>0.05 ). (3) The MBL plasma concentrations were measured in both healthy H

关 键 词：甘露糖结合凝集素 PCR-RFLP 汉族 蒙古族 突变频率 血浆含量 免疫功能 

分 类 号：R346[医药卫生—基础医学]