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Molecular Characterization of a HMW Glutenin Subunit Allele Providing Evidence for Silencing of x-type Gene on Glu-B1 一个HMW谷蛋白亚基等位基因的分子特征为Glu-B1上x型基因的沉默提供证据
Pub Date : 2006-10-01 DOI: 10.1016/S0379-4172(06)60127-3
YANG Zu-Jun , LI Guang-Rong , LIU Chang , FENG Juan , ZHOU Jian-Ping , REN Zheng-Long

Understanding the molecular structure of high-molecular-weight glutenin subunit (HMW-GS) may provide useful evidence for the study on the improvement of quality of cultivated wheat and the evolution of Glu-1 alleles. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) shows that the subunits encoded by Glu-B1 were null, named 1Bxm, in a Triticum turgidum var. dicoccoides line PI94640. Primers based on the conserved regions in wheat HMW-GS gene promoter and coding sequences were used to amplify the genomic DNA of line PI94640. The PCR products were sequenced, and the total nucleotide sequence of 3 442 bp including upstream sequence of 1 070 bp was obtained. Compared with the reported gene sequences of Glu-1Bx alleles, the promoter region of the Glu-1Bxm showed close resemblance to 1Bx7. The Glu-1Bxm coding region differs from the other Glu-1Bx alleles for a deduced mature protein with only 212 residues, and a stop codon (TAA) at 637 bp downstream from the start codon was present, which was probably responsible for the silencing of x-type subunit genes at the Glu-B1 locus. Phylogenetic tree based on the nucleotide sequence alignment of HMW glutenin subunit genes showed that 1Bxm was the most ancient type of Glu-B1 alleles, suggesting that the evolution rates are different among Glu-1Bx genes. Further study on the contribution of the unique silenced Glu-B1 alleles to quality improvement was also discussed.

了解高分子量谷蛋白亚基(hwh - gs)的分子结构可以为研究栽培小麦品质的改良和谷蛋白1等位基因的进化提供有益的依据。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)结果表明,在一株小麦(Triticum turgidum var. dicoccoides) PI94640中,Glu-B1编码的亚基为零,命名为1Bxm。以小麦HMW-GS基因启动子保守区和编码序列为引物,扩增小麦PI94640的基因组DNA。对PCR产物进行测序,得到总核苷酸序列为3 442 bp,其中上游序列为1 070 bp。与已报道的Glu-1Bx等位基因序列比较,Glu-1Bxm的启动子区域与1Bx7非常相似。与其他Glu-1Bxm等位基因不同的是,Glu-1Bxm编码区只有212个残基,并且在起始密码子下游637 bp处存在一个停止密码子(TAA),这可能是导致Glu-B1位点x型亚基基因沉默的原因。基于HMW谷蛋白亚基基因核苷酸序列比对的系统发育树显示,1Bxm是最古老的Glu-B1等位基因类型,表明Glu-1Bx基因之间的进化速率存在差异。进一步探讨了独特的Glu-B1沉默等位基因对品质改善的贡献。
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引用次数: 10
Effect of Genetic Variations of the POU1F1 Gene on Growth Traits of Nanyang Cattle POU1F1基因遗传变异对南阳牛生长性状的影响
Pub Date : 2006-10-01 DOI: 10.1016/S0379-4172(06)60124-8
XUE Kai , CHEN Hong , WANG Shan , CAI Xin , LIU Bo , ZHANG Cun-Fang , LEI Chu-Zhao , WANG Xin-Zhuang , WANG Yi-Min , NIU Hui

PCR-RFLP was applied to analyze the effect of the genetic variations of the POU1F1 gene on growth traits of 100 Nanyang cattle. The results showed that the 451 bp PCR product digested with Hinf I demonstrated polymorphism in the population, which was at Hardy-Weinberg equilibrium. Moreover, the frequencies of alleles A/B in the Nanyang population were 0.465/0.535. The association of the variations of the POU1F1 gene with the growth traits in the population was analyzed. The following parameters were greater in individuals with a genotype BB than in those with an genotype AB: birth weight, average weight increase before ablactation, body height at 12 months, body weight, body length, and chest girth at 6 months and 12 months (P<0.05). The body weight at 12 months was higher in the BB individuals than in the AA individuals (P <0.05). The body weight and body sizes also showed a trend of allele B> allele A in the other age groups. Therefore, the genotype BB maybe a dominant genotype and the allele B may be a dominant allele. These results imply that the allele B of the POU1F1 gene is likely to positively affect the growth traits.

采用PCR-RFLP方法,分析了POU1F1基因的遗传变异对100头南阳牛生长性状的影响。结果表明,用Hinf I酶切的451 bp PCR产物在群体中表现出多态性,符合Hardy-Weinberg平衡。南阳人群A/B等位基因频率为0.465/0.535。分析了POU1F1基因变异与群体生长性状的关系。BB基因型个体的出生体重、断奶前平均增重、12月龄体高、6月龄和12月龄体质量、体长和胸围均高于AB基因型个体(P<0.05)。BB组12月龄体重高于AA组(P <0.05)。体重和体型也呈现出等位基因B>的变化趋势;等位基因A在其他年龄组中。因此,基因型BB可能是显性基因型,等位基因B可能是显性等位基因。这些结果表明,POU1F1基因的等位基因B可能对生长性状有积极的影响。
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引用次数: 44
Cloning and Characterization of Four B-hordein Genes from Tibetan Hull-less Barley (Hordeum vulgare subsp. vulgare) 西藏无壳大麦(Hordeum vulgare subsp) 4个b -苦苷基因的克隆与特性分析。)
Pub Date : 2006-10-01 DOI: 10.1016/S0379-4172(06)60128-5
HAN Zhao-Xue , QIAN Gang , PAN Zhi-Fen , DENG Guang-Bing , WU Fang , TANG Ya-Wei , QIANG Xiao-Lin , YU Mao-Qun

Four B-hordein genes, designated BH1-BH4, were cloned using PCR amplification from two hull-less barley cultivars, ZQ7239 and ZQ148, collected from Tibet. The results of sequencing indicated that BH1-BH4 contained complete open reading frames (ORFs). Comparison of their predicted polypeptide sequences with the published sequences suggested that they all share the same basic protein structure. Phylogenetic analysis indicated that the deduced amino-acid sequences of BH1-BH4 genes were more closely related to B-hordeins from cultivated barley (Hordeum vulgare L.) than to any other prolamins from wild barley and Aegilops tauschii. Comparison of the coding regions of BH1-BH4 genes showed that BH1 had a lower sequence identity to other previously published B-hordeins than the other three B-hordeins obtained in this study. BH1 was then cloned in a bacterial expression vector based on bacteriophage T7 RNA polymerase. The resulting plasmid produced a 28.15 kDa protein in Escherichia coli. The potential value of B-hordein genes in grain quality improvement of hull-less barley has been discussed.

从西藏无壳大麦品种ZQ7239和ZQ148中克隆了4个B-hordein基因,编号为BH1-BH4。测序结果显示BH1-BH4包含完整的开放阅读框(orf)。他们预测的多肽序列与已发表的序列的比较表明,它们都具有相同的基本蛋白质结构。系统发育分析表明,BH1-BH4基因的氨基酸序列与栽培大麦(Hordeum vulgare L.)中b -蛋白的亲缘关系比与野生大麦和黄花青稞中其他蛋白的亲缘关系更近。BH1- bh4基因编码区比较表明,与本研究获得的其他3个b -hordein相比,BH1与其他已发表的b -hordein序列同源性较低。然后将BH1克隆到基于噬菌体T7 RNA聚合酶的细菌表达载体上。质粒在大肠杆菌中产生28.15 kDa的蛋白。探讨了B-hordein基因在无壳大麦籽粒品质改良中的潜在价值。
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引用次数: 3
Structure of the Mitochondrial DNA Control Region of the Sinipercine Fishes and Their Phylogenetic Relationship 鳜类鱼类线粒体DNA控制区结构及其系统发育关系
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60112-1
ZHAO Jin-Liang, WANG Wei-Wei, LI Si-Fa, CAI Wan-Qi

The mitochondrial DNA control region of Siniperca chuatsi, S. kneri, S. scherzeri, S. obscura, S. undulata, Coreosiniperca roulei and Coreoperca whiteheadi were amplified by PCR amplification and directly sequenced. The mtDNA control region of the sinipercine fishes could be separated into three domains, namely, the terminal associated sequence domain, the central conserved sequence domain and the conserved sequence block domain. The extended terminal associated sequence (ETAS), three conserved sequence blocks (CSB-F, CSB-E, CSB-D) in the central conserved sequence domain and three conserved sequence blocks (CSB1, CSB2, CSB3) in the conserved sequence block domain were also identified. The phylogenetic relationships among these sinipercine fishes were constructed through neighbor-joining and maximum parsimony methods using Percidae and Serranidae as outgroups. Results showed that sinipercine fishes were a monophyletic group, with Siniperca forming one group, and Coreoperca forming another group. Coreosiniperca roulei did not form an independent group but was merged into the genus Siniperca. Thus it should be renamed as Siniperca roulei.

采用PCR扩增方法,对翘嘴鳜、克氏鳜、雪氏鳜、暗箱鳜、波状鳜、roulei Coreosiniperca roulei和whiteheadcoreoperca的线粒体DNA控制区进行扩增,并直接测序。鳜类鱼类mtDNA控制区可划分为末端相关序列域、中心保守序列域和保守序列块域三个结构域。鉴定出了扩展末端关联序列(ETAS)、中部保守序列域3个保守序列区(CSB-F、CSB-E、CSB-D)和中部保守序列区3个保守序列区(CSB1、CSB2、CSB3)。以鲈科(peridae)和Serranidae为外类群,通过邻居连接法和最大简约法构建了这些鲈科鱼类之间的系统发育关系。结果表明:sininiperca属单系,Coreoperca属单系。Coreosiniperca roulei没有形成一个独立的类群,而是被合并到sininiperca属中。因此它应该被重新命名为sininiperca roulei。
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引用次数: 48
RFLP and AFLP Analysis of Inter- and Intraspecific Variation of Brassica rapa and B. napus Shows that B. rapa Is an Important Genetic Resource for B. napus Improvement 油菜和甘蓝型油菜种间和种内变异的RFLP和AFLP分析表明,甘蓝型油菜是甘蓝型油菜改良的重要遗传资源
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60115-7
LIU Ren-Hu , MENG Jin-Ling

Fingerprinting of 29 accessions of oilseed rape, including seven accessions of Brassica napus, and 22 accessions of B. rapa (B. campestris) from Europe, North America, and China was analyzed using RFLP and AFLP markers. In total, 1 477 polymorphic RFLP bands and 183 polymorphic AFLP bands from 166 enzyme-probe combinations and two pairs of AFLP primers, respectively, were scored for the 29 accessions. On average, RFLP analysis showed that the Arabidopsis EST probe detected more polymorphic bands in Brassica than the random genomic probe performed. More polymorphic RFLP markers were detected with the digestion of EcoR I or BamH I than HindIII. According to the number of bands amplified from each accession, the copy numbers of each gene in the genomes of B. rapa and B. napus were estimated. The average copy numbers in B. rapa of China, B. rapa of Europe, and B. napus, were 3.2, 3.1, and 2.9, respectively. Genetic distance based on the AFLP data was well correlated with that based on the RFLP data (r = 0.72, P<0.001), but 0.39 smaller on average. Genetic diversity analysis showed that Chinese B. rapa was more polymorphic than Chinese B. napus and European materials. Some European B. napus accessions were clustered into European B. rapa, which were distinctly different from Chinese B. napus. The larger variations of Chinese accessions of B. rapa suggest that they are valuable in oilseed rape breeding. Novel strategies to use intersubgenomic heterosis between genome of B. rapa (ArAr) and genome of B. napus (AnAnCnCn) were elucidated.

利用RFLP和AFLP对来自欧洲、北美和中国的29份油菜(包括7份甘蓝型油菜和22份油菜)的指纹图谱进行了分析。从166个酶探针组合和2对AFLP引物中分别获得1 477个多态性RFLP条带和183个多态性AFLP条带。平均而言,RFLP分析显示,拟南芥EST探针比随机基因组探针检测到更多的芸苔属多态性条带。与HindIII相比,EcoR I或BamH I酶切检测到更多的多态性RFLP标记。根据每次扩增的条带数,估计了油菜和甘蓝型油菜基因组中各基因的拷贝数。中国、欧洲和甘蓝型油菜的平均拷贝数分别为3.2、3.1和2.9。基于AFLP数据的遗传距离与基于RFLP数据的遗传距离具有良好的相关性(r = 0.72, P<0.001),但平均小0.39。遗传多样性分析表明,中国油菜比中国甘蓝型油菜和欧洲油菜更具多态性。部分欧洲甘蓝型油菜品种聚为欧洲甘蓝型油菜,与中国甘蓝型油菜品种有明显差异。中国油菜品种变异较大,说明其在油菜育种中具有一定的应用价值。提出了利用油菜B. rapa (ArAr)和油菜B. napus (AnAnCnCn)基因组亚基因组间杂种优势的新策略。
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引用次数: 12
Construction of Fingerprinting and Genetic Diversity of Mulberry Cultivars in China by ISSR Markers 利用ISSR标记构建中国桑树品种指纹图谱及遗传多样性
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60119-4
ZHAO Wei-Guo , MIAO Xue-Xia , ZANG Bo , ZHANG Lin , PAN Yi-Le , HUANG Yong-Ping

The ISSR fingerprintings of 24 mulberry cultivars were constructed. Totally 80 bands were produced using 17 primers selected from 20 primers. Of them, 40 bands showed polymorphism. From the bands amplified, there were three independent ways to identify the mulberry varieties, such as unique ISSR markers, unique band patterns and a combination of the band patterns provided by different primers. ISSRs were very effective in differentiating the mulberry varieties. The mean genetic similarity coefficient, the mean Nei's gene diversity (h), and the mean Shannon's Information index (I) of mulberry cultivars were 0.8731, 0.1210, and 0.1942, respectively. This suggests that the genetic diversity of mulberry cultivars was low and the genetic base was narrow. Both UPGMA cluster and PCA (Principal Coordinates Analysis) analysis showed clear genetic relationships among the 24 mulberry cultivars. The major clusters were related to known pedigree relationships.

构建了24个桑树品种的ISSR指纹图谱。从20条引物中筛选出17条引物,共得到80条条带。其中有40个条带存在多态性。从扩增的条带来看,桑树品种的鉴定有三种独立的方法,即独特的ISSR标记、独特的条带模式和不同引物提供的条带模式组合。ISSRs对桑树品种的鉴别非常有效。桑树品种遗传相似系数的平均值为0.8731,Nei’s基因多样性的平均值为0.1210,Shannon’s信息指数的平均值为0.1942。这说明桑树品种遗传多样性较低,遗传基础较窄。UPGMA聚类分析和主坐标分析结果表明,24个桑树品种间存在明显的亲缘关系。主要的群集与已知的系谱关系有关。
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引用次数: 12
Linkage and Association Between CA Repeat Polymorphism of the TNFR2 Gene and Obesity Phenotypes in Two Independent Caucasian Populations 两个独立高加索人群中TNFR2基因CA重复多态性与肥胖表型的连锁和关联
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60110-8
HUANG Qing-Yang , SHEN Hui , DENG Hong-Yi , Theresa Conway , Leo Elze , K. Michael Davies , Robert R. Recker , DENG Hong-Wen

Previously, our group has reported a suggestive linkage evidence of 1p36 with body mass index (BMI) (LOD = 2.09). The tumor necrosis factor receptor 2 (TNFR2) at 1p36 is an excellent positional and functional candidate gene for obesity. In this study, we have investigated the linkage and association between the TNFR2 gene and obesity phenotypes in two large independent samples, using the quantitative transmission disequilibrium tests (QTDT). The first group was made up of 1 836 individuals from 79 multi-generation pedigrees. The second group was a randomly ascertained set of 636 individuals from 157 US Caucasian nuclear families. Obesity phenotypes tested include BMI, fat mass, and percentage fat mass (PFM). A significant result (P = 0.0056) was observed for linkage with BMI in the sample of the multigenerational pedigrees. Our data support the TNFR2 gene as a quantitative trait locus (QTL) underlying BMI variation in the Caucasian populations.

本课题组此前报道了1p36与身体质量指数(BMI) (LOD = 2.09)的关联证据。肿瘤坏死因子受体2 (TNFR2)位于1p36,是一个很好的定位和功能的肥胖候选基因。在这项研究中,我们使用定量传递不平衡测试(QTDT)在两个大型独立样本中研究了TNFR2基因与肥胖表型之间的联系和关联。第一组由来自79个多代谱系的1836个个体组成。第二组是随机确定的来自157个美国高加索核心家庭的636个人。肥胖表型测试包括BMI、脂肪量和脂肪量百分比(PFM)。在多代谱系样本中,观察到与BMI的关联显著结果(P = 0.0056)。我们的数据支持TNFR2基因作为高加索人群BMI变异的数量性状位点(QTL)。
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引用次数: 1
The Tissue Distribution and Developmental Changes of ghrelin mRNA Expression in Sheep 绵羊胃饥饿素mRNA表达的组织分布及发育变化
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60114-5
HUANG Zhi-Guo , XIONG Li , LIU Zhen-Shan , QIAO Yong , DAI Rong , XIE Zhuang , LIU Shou-Ren , SHI Guo-Qing , LIU Guo-Qing

Male Kazak sheep and Xinjiang fine wool sheep, six for each different age group (days 2, 30, 60, 90 and 120), were used in the present study to investigate the tissue distribution and developmental changes of ghrelin mRNA expression in abomasum; however, there was no 120-day-old Kazak sheep. After measurement of body weight, the tissues such as hypothalamus, pituitary, heart, liver, rumen, reticulum, omasum, abomasum, duodenum, and longissimus dorsi muscle were sampled. And the total RNA of different tissues was extracted to determine the abundance of ghrelin mRNA by RT-PCR and real-time PCR. The results showed that (1) for both breeds, body weight among different ages was significantly different (P<0.05). And from day 30 to 90, the body weight of Kazak was significantly higher than that of Xinjiang (P<0.01); (2) Ghrelin mRNA existed in all the above tissues and was significantly higher in the abomasum than in other tissues (P<0.05); (3) the temporal patterns of abomasum ghrelin mRNA expression in Kazak and Xinjiang were similar. From day 2 to 60 in Kazak and 2 to 90 in Xinjiang, there was a steady increase in the ghrelin mRNA level. By day 60 in Kazak and day 90 in Xinjiang, the level reached a plateau and remained steady. These results also demonstrated that from birth to day 90, ghrelin mRNA level was significantly higher in Kazak than in Xinjiang (P<0.01).

选取哈萨克羊和新疆细毛羊,每组各6只(第2、30、60、90和120天),研究胃饥饿素mRNA在皱胃组织分布和发育变化;然而,没有120日龄的哈萨克羊。测量体重后,取下丘脑、垂体、心脏、肝脏、瘤胃、网状、瓣胃、皱胃、十二指肠、背最长肌等组织标本。提取不同组织的总RNA,采用RT-PCR和real-time PCR检测ghrelin mRNA的丰度。结果表明:(1)两个品种不同年龄间体重差异显著(P<0.05)。第30 ~ 90天,哈萨克猪的体重极显著高于新疆猪(p < 0.01);(2)胃饥饿素mRNA均存在于上述组织中,且在皱胃中含量显著高于其他组织(P<0.05);(3)哈萨克族和新疆族皱胃胃饥饿素mRNA表达的时间格局相似。哈萨克和新疆分别在第2 ~ 60天和第2 ~ 90天,胃饥饿素mRNA水平稳步升高。哈萨克和新疆分别在第60天和第90天达到高原水平并保持稳定。这些结果还表明,从出生到第90天,哈萨克族的ghrelin mRNA水平显著高于新疆族(P<0.01)。
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引用次数: 21
Progress in Plant CACTA Elements 植物CACTA元件研究进展
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60109-1
TIAN Ping-Fang

Transposable elements are DNA fragments that can insert new chromosomal locations. On the basis of the mechanism of transposition, transposable elements were divided into two classes. Class 1 elements were retroelements that used reverse transposase to transpose by an RNA intermediate. Class 2 elements or DNA transposons transposed directly from DNA to DNA. Of the Class 2 elements, CACTA superfamily, so far identified exclusively in plants and previously regarded as low-copy-transposon for the conserved mechanism of propagation, recently received considerable interest because of their increasing evidence reiterating their high copies in some plant genomes. This article aimed at outlining CACTA elements with regard to their structure, transposition, and utilization.

转座因子是可以插入新的染色体位置的DNA片段。根据转座机理,将可转座因子分为两类。第1类元件是逆转录元件,使用反转座酶通过RNA中间体进行转座。第2类元件或DNA转座子直接从DNA转到DNA。在2类元件中,CACTA超家族迄今为止仅在植物中发现,以前被认为是保守繁殖机制的低拷贝转座子,最近由于越来越多的证据重申其在某些植物基因组中的高拷贝而引起了相当大的兴趣。本文旨在概述CACTA要素的结构、转换和利用。
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引用次数: 13
Salt Induces Expression of RH3.2A, Encoding an H3.2-type Histone H3 Protein in Rice (Oryza sativa L.) 盐诱导水稻h3.2型组蛋白H3蛋白RH3.2A的表达
Pub Date : 2006-09-01 DOI: 10.1016/S0379-4172(06)60117-0
QIU Sheng-Ping, HUANG Ji, PAN Li-Juan, WANG Mei-Mei, ZHANG Hong-Sheng

Histone H3 is one of the four histones, along with H2A, H2B, and H4, which form the eukaryotic nucleosome octamer core. In this study, a new gene RH3.2A encoding an H3.2-type histone H3 protein from rice (Oryza sativa L.) was reported. RH3.2A was cloned through RT-PCR from salt-treated rice seedlings. This gene encoded a protein of 136 amino acid residues that were similar to some plant histone H3 proteins reported previously. However, the cDNA sequence of RH3.2A and other rice H3 genes were different. Alignment of RH3.2A encoding protein with other plant histone H3 proteins revealed that three amino acid residues (32, 88, and 91) were markedly different between H3.1-type and H3.2-type proteins. The mRNA expression analysis of RH3.2A revealed that RH3.2A gene was upregulated by salt stress in rice roots and ABA treatment in seedlings. The potential role of RH3.2A during salt stress was discussed.

组蛋白H3是与H2A、H2B和H4一起构成真核小体八聚体核心的四种组蛋白之一。本研究报道了一个从水稻(Oryza sativa L.)中编码h3.2型组蛋白H3蛋白的新基因RH3.2A。通过RT-PCR从盐处理水稻幼苗中克隆RH3.2A。该基因编码了一个含有136个氨基酸残基的蛋白,与之前报道的一些植物组蛋白H3蛋白相似。然而,RH3.2A与其他水稻H3基因的cDNA序列不同。将RH3.2A编码蛋白与其他植物组蛋白H3蛋白比对发现,h3.1型和h3.2型蛋白的3个氨基酸残基(32、88和91)存在显著差异。通过对RH3.2A mRNA的表达分析发现,水稻根系盐胁迫和幼苗ABA处理导致RH3.2A基因表达上调。讨论了RH3.2A在盐胁迫中的潜在作用。
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引用次数: 6
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Acta Genetica Sinica
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