Frequent mtDNA mutations and its role in gastric carcinogenesis

In order to disclose the relationship between mutations of mitochondrial DNA (mtDNA) and gastric carcinogenesis, we screened the entire mtDNA sequence in 30 cases of human gastric cancer and matched normal tissues by using denaturing high-performance liquid chromatography (DHPLC) and DNA sequencing. Our data showed that high frequency (66.7%, 20/30) of mitochondrial genome mutation occurred in gastric cancer. Among these variants, 17 cases (56.7%, 17/30) were identified to be somatic mutation. High level mutant frequency was found in ND4, ND5 coding genes and D-loop control region, which was 36.7%, 26.7% and 30% respectively. Comparing with complexes Ⅲ, Ⅳ and Ⅴof the electron transport chain, we found that variants appeared to be more frequent in the subunit genes of complexⅠ. Most of mutations were base substitutions (85.4%, 41/48). Our results suggested that mutations of subunit genes encoding complexⅠ, especially ND3, ND4 and ND5 genes, might contribute to human gastric carcinogenesis.     

裂足轮虫还是裂足臂尾轮虫

通过分析比较了裂足轮虫(Brachionus diuersicornis)、方型臂尾轮虫(B．quadridentatus)、壶状臂尾轮虫(B．urceus)、矩形臂尾轮虫(B．1eydigi)、萼花臂尾轮虫(B．calyciflorus)等5种轮虫的线粒体细胞色素氧化酶亚基I(COI）的部分序列，并结合4种海水臂尾轮虫的序列数据，用西氏晶囊轮虫(Asplanchna．sieboldi)作外群构建UPGMA树和NJ树，探讨了烈足轮虫的分类学问题，认为将裂足轮虫归属千臂尾轮虫属市为合适。     

检测线粒体DNA SNPs的快速测定法--引物延伸-飞行时间质谱法

用PCR扩增出人类mtDNA HV Ⅰ区443bp片段作为模板,以nt16093位点特异的引物进行引物延伸反应。产物经纯化后利用MALDI-TOF质谱技术进行检测,从而建立起引物延伸—飞行时间质谱法快速检测SNPs的技术,并用该法对人类mtDNA HV Ⅰ区SNPs位点nt16093的T／C多态性进行频率调查。结果为广州地区汉族人群的mtDNA HV Ⅰ区多态位点nt16093T型的频率为89．6％,C型的频率为10．4％。并且发现3例异质性。     

Pitfalls in the analysis of ancient human mtDNA

The retrieval of DNA from ancient human specimens is not always successful owing to DNA deterioration and contamination although it is vital to provide new insights into the genetic structure of ancient people and to reconstruct the past history. Normally, only short DNA fragments can be retrieved from the ancient specimens. How to identify the authenticity of DNA obtained and to uncover the information it contained are difficult. We employed the ancient mtDNAs reported from Central Asia (including Xinjiang, China) as an example to discern potentially extraneous DNA contamination based on the updated mtDNA phylogeny derived from mtDNA control region, coding region, as well as complete sequence information. Our results demonstrated that many mtDNAs reported are more or less problematic.Startim, from a reliable mtDNA phylogeney and combining the available modern data into analysis, one can ascertain the authenticity of the ancient DNA, distinguish the potential errors in a data set, and efficiently decipher the meager information it harbored. The reappraisal of the mtDNAs with the age of more than 2000 years from Central Asia gave support to the suggestion of extensively (pre)historical gene admixture in this region.     

大熊猫线粒体DNA酶切片段长度多态性研究

用１２种限制性内切酶（ＢａｍＨⅠ，ＥｃｏＲⅠ，ＥｃｏＲⅤ，ＸｈｏⅠ，ＰｓｔⅠ，ＭｓｐⅠ，ＸｂａⅠ，ＰｖｕⅡ，ＨｉｎｄⅢ，ＨａｅⅢ，ＳａｃⅠ，ＨｐａⅡ）对秦岭大熊猫线粒体ＤＮＡ进行了酶切。测定和分析了每种酶所产生片段的数量和大小，并与四川大熊猫的限制性片段进行了比较，发现在相同的６种酶的１６个酶切位点中有一个不同，表明这两地群体间存在着线粒体ＤＮＡ的多态性。其研究结果为进一步研究和保护大熊猫提供了重要依据。     

Single nucleotide polymorphisms of mitochondrial DNA HVS‐I and HVS‐II in Chinese Bai ethnic group

For forensic and population genetic purposes, a total of 125 unrelated volunteers’ blood samples were collected from Chinese Bai ethnic minority group to analyze sequence variation of two hypervariable segments (HVS‐I and HVS‐II) in the mitochondrial DNA control region. Comparing the HVS‐I and HVS‐II sequences of the 125 Chinese Bais to the Anderson reference sequence, we found 86 polymorphic loci in HVS‐I and 40 in HVS‐II in mitochondrial DNA sequences of the Chinese Bai ethnic minority group, which defined 93 and 53 different haplotypes, respectively. Haplotype diversity and the mean pairwise differences were 0.992 ± 0.003 and 6.553 in HVS‐I, and 0.877 ± 0.027 and 2.407 in HVS‐II, respectively. We defined four macrohaplogroups R, M, N and D with the proportions ranging from 9.6% to 40.0%. With the analysis of the hypervariable domain from nucleotide 16 180–16 193 in HVS‐I, our study revealed new haplotypes of sequence variations. In addition, the Fst metric, phylogenetic tree, and principal component analysis demonstrated a close genetic relationship between the Bai group and Chinese Han populations from South China, Changsha, and Guangdong. The results support that the Bai group is a multiorigin ethnic minority that has merged with the Chinese Han population.     

MGB probe assay for rapid detection of mtDNAl1778 mutation in the Chinese LHON patients by real-time PCR

Objective： Leber＇s hereditary optic neuropathy （LHON） is a maternally inherited degeneration of the optic nerve caused by point mutations of mitochondrial DNA （mtDNA）. Many unsolved questions regarding the penetrance and pathophysiological mechanism of LHON demand efficient and reliable mutation testing. This study aims to develop a minor groove binder （MGB） probe assay for rapid detection of mtDNA11778 mutation and heteroplasmy in Chinese LHON patients by real-time polymerase chain reaction （PCR）. Methods： Forty-eight patients suspected of having LHON and their maternal relatives underwent a molecular genetic evaluation, with 20 normal individuals as a control group at the same time. A real-time PCR involving two MGB probes was used to detect the mtDNA 1 1778 mutation and heteroplasmy. A linear standard curve was obtained by pUCmLHONG and pUCmLHONA clones. Results： All 48 LHON patients and their maternal relatives were positive for rntDNA11778 mutation in our assay, 27 heteroplasmic and 21 homoplasmic. Eighteen cases did not show an occurrence of the disease, while 9 developed the disease among the 27 heteroplasmic mutation cases. Eleven did not show an occurrence of the disease, while 10 cases developed the disease among 21 homoplasmic mutation cases. There was a significant difference in the incidence between the heteroplasmic and the homoplasmic mutation types. The time needed for running a real-time PCR assay was only 80 min. Conclusion： This real-time PCR assay is a rapid, reliable method for mtDNA mutation detection as well as heteroplasmy quantification. Detecting this ratio is very important for predicting phenotypic expression of unaffected carriers.     

中国部分地方牛种mtDNA D-loop区全序列的遗传多样性与系统进化分析

利用PCR测序及生物信息学分析技术,对我国5个地方黄牛品种、5个地方水牛品种及2个地方牦牛品种的mtDNA D-loop区全序列进行PCR扩增以及核苷酸多样度、单倍型多样度分析,发现中国地方黄牛、水牛与牦牛具有丰富的遗传多样性.对试验牛mtDNA D-loop区全序列与牛亚科代表性物种黄牛、水牛、家牦牛、野牦牛、欧洲普通牛、印度瘤牛以及摩拉水牛相应序列进行系统发育分析.结果显示:黄牛与牦牛的亲缘关系较近,它们与水牛的亲缘关系较远;中国水牛属于沼泽型水牛,也有少量江河型水牛渐渗入中国水牛群体;中国黄牛为普通牛和瘤牛的混合母系起源;进化树显示高原牦牛与野牦牛的亲缘关系较近,环湖牦牛与家牦牛的亲缘关系较近.     

4个群体西太公鱼mtDNAD—loop区段的限制性片段长度多态性分析

应用PCR技术对天津市于桥水库、北京市密云水库、日本北海道和日本青森县4个群体共151尾西太公鱼（Hypomesusnipponensis）的mtDNAD-loop区段进行扩增,得到长度约为1．8kbp的扩增片段．扩增出的DNA片段使用13种核酸内切限制酶进行酶切,其中8种核酸内切限制酶（AluI、DraI、HaeⅢ、Hindm、HinfI、MboI、TaqI和XspI）有酶切位点,5种核酸内切限制酶（AluI、HinfI、MboI、TaqI和XspI）个体间存在变异．不同酶切结果组合后,共得到28种单倍型,于桥水库、密云水库、北海道和青森西太公鱼均以单倍型6为主,分别为36．36％、38．30％、43．33％和26．67％;于桥水库、密云水库、北海道及青森西太公鱼单倍型多样性指数（危）分别为0．8171、0．8141、0．7793和0．8828;核苷酸多样性指数Or）分别为0．007918、0．007326、0．007255和0．009710．于桥水库西太公鱼与密云水库西太公鱼之间的Rogers遗传距离最小,为0．1499;于桥水库西太公鱼与日本青森西太公鱼之间的Rogers遗传距离最大,为0．2215;日本北海道西太公鱼与密云水库西太公鱼之间的遗传距离为0．1511,大于于桥水库西太公鱼与密云西太公鱼之间的遗传距离,但小于日本境内北海道西太公鱼与青森西太公鱼之间的遗传距离．     

中国白兔线粒体DNA限制性图谱

用 ６ 种限制性内切酶分析了中国白兔的线粒体 Ｄ Ｎ Ａ（ｍ ｔ Ｄ Ｎ Ａ）。测得其相对分子质量约为１６．８, Ｅｃｏ ＲⅤ, Ｂａｍ ＨⅠ, ＰｓｔⅠ, Ｅｃｏ ＲⅠ, ＨｉｎｄⅢ在中国白兔 ｍ ｔ Ｄ Ｎ Ａ 上分别有 １,２,２,２,６ 个切点, ＳａｌⅠ 在其上没有切点。根据单酶降解和双酶降解片段的相对分子质量,构建了中国白兔ｍ ｔ Ｄ Ｎ Ａ的限制性内切酶图谱。