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[Identification of differential genomic genes of Mycobacterium tuberculosis H37Rv and attenuated strain H37Ra by suppression subtractive hybridization]. [抑制减法杂交鉴定结核分枝杆菌H37Rv与减毒菌株H37Ra的差异基因组基因]。
Zhi-Hong Xiong, Yu-Hui Zhuang, Guo-Li Li

To study the virulence-related genes in Mycobacterium tuberculosis, we used suppression subtractive hybridization to clone the differential genomic genes between Mycobacterium tuberculosis virulence strain H37Rv and attenuated strain H37Ra. All of 54 different genes were cloned, sequenced and analyzed by Southern-blotting. Two different DNA fragments in H37Ra are new genes so far, and get the new Genbank number AY534505 and AY560011. Eight different DNA fragments in H37Rv were obtained. One is the fragment of a gene coding virulence factor mce; one fragment belongs to the gene coding for purC synthenzyme; one for PE family protein; the other 4 fragments for putative gene; and the last one is a non-coding fragment. PCR analysis indicated that 2 of the different genes were present exclusively in the clinical virulent strain and in H37Rv, but not in the clinical avirulent strain and in H37Ra. The novel differential genes may provide an important clue for studying the mechanism of M. tuberculosis pathogenesis.

为了研究结核分枝杆菌毒力相关基因,我们采用抑制减法杂交技术克隆结核分枝杆菌毒力菌株H37Rv与减毒菌株H37Ra的差异基因组基因。对54个不同基因进行克隆、测序和Southern-blotting分析。H37Ra中两个不同的DNA片段是迄今为止的新基因,并获得新的基因库编号AY534505和AY560011。在H37Rv中获得了8个不同的DNA片段。一个是编码毒力因子mce的基因片段;一个片段属于编码purC合成酶的基因;一个为PE家族蛋白;其余4个片段为推定基因;最后一个是非编码片段。PCR分析表明,其中2个基因只存在于临床毒株和H37Rv中,而不存在于临床无毒毒株和H37Ra中。这些新的差异基因可能为研究结核分枝杆菌的发病机制提供重要线索。
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引用次数: 0
[Establishment of transgenic mouse line skin-specifically expressing hCTLA4-Ig]. [建立皮肤特异性表达hCTLA4-Ig的转基因小鼠系]。
Yong Wang, Feng-Chao Wang, Hong Wei, Yong Ni, Jun Wu, Xiang Gao

hCTLA4-Ig,which is capable of inhibiting T cell activation and rendering T cell to be anergy by blocking co-stimulatory signal pathway, is recognized as a potential therapeutic molecule to extend the survival of skin graft in burn medicine. To investigate whether the survival of skin graft can be further extended by skin-specific expression of hCTLA4-Ig, a skin-specific expression vector was constructed by pacing the encoding sequence of hCTLA4-Ig under the drive of K14 promoter, and a transgenic mouse line was established with the vector. RT-PCR and Northern blot assay indicated that hCTLA4-Ig was skin-specifically expressed in transgenic mice at a rather high level. Compared with the expression of GAPDH, the expression of hCTLA4-Ig remained constant through generations and over lifetime. The results of this paper indicated that a transgenic mouse line skin-specifically and constitutively expressing hCTLA4-Ig had been established.

hCTLA4-Ig能够通过阻断共刺激信号通路抑制T细胞活化,使T细胞处于能量状态,是烧伤医学中公认的延长植皮存活的潜在治疗分子。为了研究皮肤特异性表达hCTLA4-Ig是否能进一步延长移植物的存活时间,在K14启动子的驱动下,通过调整hCTLA4-Ig的编码序列构建皮肤特异性表达载体,并用该载体建立转基因小鼠系。RT-PCR和Northern blot结果表明,hCTLA4-Ig在转基因小鼠中具有较高的皮肤特异性表达水平。与GAPDH的表达相比,hCTLA4-Ig的表达在几代和一生中保持不变。结果表明,已建立了一种皮肤特异性和组成性表达hCTLA4-Ig的转基因小鼠系。
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引用次数: 0
[Methodology of mapping quantitative trait loci for discrete traits using maximum likelihood]. [使用最大似然方法为离散性状绘制数量性状位点的方法]。
Zong-Jun Yin, Qin Zhang, Hong-Quan Chen, Ji-Gang Zhang, Xiang-Dong Ding, Chun-Kao Wang

The maximum likelihood method was used to compare the efficiency of interval mapping with either the threshold model or the linear model. The irfluencing factors of quantitative trait loci (QTL) detection efficiency (e.g. QTL effect, heritability and incidence of categories) were simulated in our study. Daughter design with multiple families was applied, and the number of segregating population was 500. The results showed that the threshold model was superior in terms of parameter estimation. It was a more efficient and accurate model of QTL mapping for discrete traits. In addition, the accuracy of QTL mapping depended on the effect of the putative QTL, the value of heritability and incidence directly. With an increase of QTL effect, heritability and incidence of categories, the accuracy of QTL mapping improved correspondingly.

利用极大似然法对区间映射与阈值模型和线性模型的效率进行比较。本研究模拟了影响QTL检测效率的因素(如QTL效应、遗传力和类别发生率)。采用多家庭子代设计,隔离人口500人。结果表明,阈值模型在参数估计方面具有优越性。为离散性状的QTL定位提供了一种高效、准确的模型。此外,QTL定位的准确性直接取决于推测QTL的作用、遗传力值和发生率。随着QTL效应、遗传力和类别发生率的增加,QTL定位的准确性相应提高。
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引用次数: 0
Molecular evidence of barley yellow dwarf virus replication/movement suppressed by the resistance gene Bdv2. 抗基因Bdv2抑制大麦黄矮病毒复制/运动的分子证据
Xiao-Dong Liu, Zeng-Yan Zhang, Yan Liu, Zhi-Yong Xin

A wheat-Thinopyrum intermedium translocation line YW642 possesses the resistance to GAV serotype of barley yellow dwarf virus (BYDV), in which the resistance gene Bdv2 is derived from the chromosome 7X of Thinopyrum intermedium group 7. It is interesting to analyze BYDV accumulation content in the resistant and susceptible wheat plants for controlling BYDV disease and understanding the resistance mechanism against BYDV. In the paper, semi-quantitative reverse-transcription PCR (RT-PCR) was used to detect and quantify BYDV-GAV in the resistant and susceptible plants using specific primers for the coat protein (CP) and RNA-dependent RNA polymerase (RdRp) genes of BYDV-GAV serotype. On the inoculation site, the amount of the virus in the resistant wheat line (YW642) was much lower compared to the susceptible sib line (YW641). There was small amount of the virus could be detected in YW642 at 2-5 days post infestation (dpi), afterwards the amount of virus decreased and no virus could be detected at 14 and 16 dpi. In the uninoculated upper leaves, no BYDV was detected in YW642 from 1 to 14 dpi, while the virus could be detected at 3 dpi and then accumulated rapidly in YW641. These results showed at molecular level that the replication and/or movement of BYDV-GAV were strongly suppressed in YW642, presumably owing to the action of the BdV2 gene.

一株小麦-Thinopyrum中间易位系YW642对大麦黄矮病毒(BYDV) GAV血清型具有抗性,其中抗性基因Bdv2来源于Thinopyrum中间组7的7X染色体。分析小麦抗病和易感植株中BYDV的积累含量,对防治BYDV及了解其抗病机制具有重要意义。利用BYDV-GAV血清型外壳蛋白(CP)和RNA依赖RNA聚合酶(RdRp)基因的特异性引物,采用半定量反转录PCR (RT-PCR)技术对抗性和易感植物中的BYDV-GAV进行检测和定量。在接种现场,抗性小麦品系(YW642)的病毒量远低于易感同胞品系(YW641)。YW642侵染后2 ~ 5 d可检出少量病毒,侵染后病毒量下降,14、16 dpi时未检出病毒。在未接种的上叶中,YW642在1 ~ 14 dpi未检测到BYDV,而在3 dpi时可以检测到病毒,然后在YW641中迅速积累。这些结果表明,在分子水平上,BYDV-GAV的复制和/或运动在YW642中被强烈抑制,可能是由于BdV2基因的作用。
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引用次数: 0
[Fine localization of rice EUI1 gene controlling elongation of the uppermost internode]. 水稻最上节间伸长控制基因EUI1的精细定位。
Ren-Xiao Wang, Pei-Jin Li, Hong-Qi Chen, Shao-Kai Wen, Jia-Yang Li, Xu-Dong Zhu

The height of rice is one of the important agricultural traits, which affects on the rice architecture and production directly. The investigation of the length of the uppermost internode showed that the elongation mutation of the first internode is controlled by a recessive nuclear locus. The gene is localized between the STS marker of E30531 and the CAPS marker of C903 on the long arm of chromosome 5, apart from 6.7 cM and 2.8 cM respectively. The fine localization maps the gene between 0.3 cM flanks, which will help clone this gene and study the mechanism of the gene function.

水稻株高是重要的农业性状之一,直接影响到水稻的结构和产量。对最高节间长度的研究表明,第一节间的伸长突变受一个隐性核位点控制。该基因定位于5号染色体长臂上E30531的STS标记和C903的CAPS标记之间,距离分别为6.7 cM和2.8 cM。精细定位将基因定位在0.3 cM的两侧,这将有助于该基因的克隆和功能机制的研究。
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引用次数: 0
[Functions of FANCL in primordial germ cell formation and Fanconi anemia]. [FANCL在原始生殖细胞形成和范可尼贫血中的作用]。
Qing-Guo Zhao, Bai-Song Lu, Pei-Tang Huang

Fanconi anemia (FA) is a rare autosomal recessive disorder characterized clinically by congenital abnormalities, progressive bone marrow failure and cancer susceptibility. Cells from individuals with Fanconi anemia manifest features of spontaneous chromosomal instability and hypersensitivity to DNA cross-linking agents such as mitomycin C. Over 11 known Fanconi anemia gene products are involved in DNA damage response pathway. In the pathway, monoubiquitination of FANCD2 is a key step. A novel protein FANCL is a component of the nuclear FA complex, functioned as an ubiquitin E3 ligase and monoubiquitinylated FANCD2. FANCD2-Ub is targeted to chromatin, where it interacts with BRCA2 to repair DNA damage. In early embryo stage, FA pathway is probably involved in proliferation of PGCs. Mice deficient in FA proteins, such as FANCL, FANCC and FANCA, have a drastic reduction of primordial germ cells (PGC), resulting in male and female infertility in adult. In the adult male, FANCL and a few testis-specific proteins, GGN1 (gametogenetin protein 1), GGNBP1 (gametogenetin binding protein 1), GGNBP2 and OAZ3 (ornithine decarboxylase antizyme 3) form a novel testis-specific complex functioning in spermatogenesis. FANCL is involved in proliferation of PGCs in early embryo stage, and development of germ cells in adult.

范可尼贫血(FA)是一种罕见的常染色体隐性遗传病,临床表现为先天性异常、进行性骨髓衰竭和癌症易感性。范可尼贫血患者的细胞表现出自发性染色体不稳定和对丝裂霉素c等DNA交联剂的超敏反应,已知有11种范可尼贫血基因产物参与DNA损伤反应途径。在这一途径中,FANCD2的单泛素化是关键步骤。一种新的蛋白FANCL是核FA复合物的一个组成部分,作为泛素E3连接酶和单泛素化的FANCD2。FANCD2-Ub靶向染色质,在那里它与BRCA2相互作用以修复DNA损伤。在胚胎早期,FA通路可能参与了PGCs的增殖。缺乏FANCL、FANCC和FANCA等FA蛋白的小鼠,其原始生殖细胞(PGC)急剧减少,导致成年雄性和雌性不育。在成年雄性中,FANCL和一些睾丸特异性蛋白GGN1(配子基因蛋白1)、GGNBP1(配子基因结合蛋白1)、GGNBP2和OAZ3(鸟氨酸脱羧酶抗酶3)形成了一种新的睾丸特异性复合物,在精子发生中起作用。FANCL参与胚胎早期PGCs的增殖和成体生殖细胞的发育。
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引用次数: 0
Expression pattern and preliminary functional analysis of human CREB4 gene. 人CREB4基因表达模式及初步功能分析。
Yong-Juan Gao, Gen-Tao Cao, Gang Yin, Xin Wang, Chao-Neng Ji, Shao-Hua Gu, Xiao-Hua Ni

cAMP response element-binding (CREB) proteins are a family of mammalian transcription activators that mediate cAMP and calcium-dependent gene expression through the cAMP response element (CRE). CREB4 is a novel member of the human CREB family. RT-PCR showed CREB4 transcripts were found in lung carcinoma LX-1, colon adenocarcinoma CX-1, prastatic adenocarcinoma PC-3, colon carcinoma G1-112, and pancreatic adenocarcinoma G1-103. Constructing CREB4 and CREB(215-395aa) fusion protein with the entire prokaryotic LexA protein respectively disclosed that CREB4 protein functioned as a transcription activator and its N-terminal accounted for the activation ability. Furthermore,a fusion protein of GFP and full-length CREB4 was localized in cytoplasm,whereas the fusion protein of GFP and a deletion mutant lacking the C-terminal putative transmembrane domain was translocated in nucleus. Our results suggested that putative transmembrane domain of CREB4 protein was associated with modulation of its function for the transcriptional activation.

cAMP反应元件结合蛋白(CREB)是一个哺乳动物转录激活因子家族,通过cAMP反应元件(CRE)介导cAMP和钙依赖性基因的表达。CREB4是人类CREB家族的新成员。RT-PCR结果显示,在肺癌LX-1、结肠癌CX-1、前列腺癌PC-3、结肠癌G1-112、胰腺腺癌G1-103中均发现了CREB4转录物。分别与整个原核生物LexA蛋白构建CREB4和CREB(215-395aa)融合蛋白,发现CREB4蛋白具有转录激活因子的功能,其n端具有激活能力。此外,GFP与全长CREB4的融合蛋白定位在细胞质中,而GFP与缺乏c端跨膜结构域的缺失突变体的融合蛋白在细胞核中易位。我们的研究结果表明,CREB4蛋白的跨膜结构域与其转录激活功能的调节有关。
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引用次数: 0
[QTLs mapping and genetic analysis of tiller angle in rice (Oryza sativa L.)]. 水稻分蘖角qtl定位与遗传分析[j]。
Chuan-Yuan Yu, Yu-Qiang Liu, Ling Jiang, Chun-Ming Wang, Hu-Qu Zhai, Jian-Min Wan

Tiller angle is one of the most important morphological characters that has a significant effect on the formation of rice high-yield population. The tiller angles were measured in a japonica/indica RIL population with 71 lines and a genome-wide chromosome segment substitution line (CSSL) population with 65 lines at two experimental sites. A transgressive segregation was observed in both populations. QTL analysis of tiller angle was conducted based on the saturated RFLP marker linkage map and the CSSL graphical genetype. Five main-effect QTLs and three pairs of epstatic loci were detected in the RIL population. A main QTL, qTA-9, located on chromosome 9 at XNpb108 - C506, was identified at both experiment sites. The positive allele TA-9(I) on qTA-9 was contributed by indica rice IR24 with 28.6% average contribution to variance. Meanwhile, an analysis of CSSL graphical genetypes also showed that there was a positive allele on the IR24 chromosome substitution segment delimited by RFLP marker C609 and C506 with approximate 15 cM interval, which proved the existence of qTA-9. The TA-9(I) could increase tiller angle by about 15 degrees in japonica Asominori background under the two environments. The measurement of the F1 from the cross between background parent and CSSL AIS68 with TA-9(I) and the analysis of F2 population indicated that the TA-9(I) was an incomplete dominant gene. Genetype x environment interaction(G xE) was not widely present except a pair of epistatic loci with 5.32% contribution to variance of tiller angles and a relatively small additive effect. The combining action of the additive effect of the genes from both parents and the two-loci epistasis-effect may be responsible for the transgressive segregation of tiller angle in rice population. The value and approach of application of TA-9(I) in hybrid rice breeding program were discussed.

分蘖角是影响水稻高产群体形成的重要形态性状之一。在两个试验点分别测定了71个粳稻/籼稻RIL群体和65个全基因组染色体片段代换系(CSSL)群体的分蘖角。在两个种群中均观察到海侵隔离现象。利用饱和RFLP标记连锁图谱和CSSL图形基因型对分蘖角进行QTL分析。在RIL群体中检测到5个主效qtl和3对易感位点。在XNpb108 - C506的9号染色体上发现了一个主QTL qTA-9。qTA-9上的正等位基因TA-9(I)由籼稻IR24贡献,平均方差贡献率为28.6%。同时,对CSSL图形基因型分析也发现,在RFLP标记C609和C506所分隔的IR24染色体代换片段上,间隔约为15 cM,存在一个阳性等位基因,证明qTA-9的存在。在两种环境下,TA-9(I)均能使小野粳稻的分蘖角增加约15度。本底亲本与CSSL AIS68杂交TA-9(I)的F1测定和F2群体分析表明TA-9(I)为不完全显性基因。基因型x环境互作(gxe)除对分蘖角变异贡献率为5.32%,加性效应较小外,其他基因型x环境互作(gxe)并不广泛存在。两亲本基因的加性效应和双位点上位效应的共同作用可能是水稻群体分蘖角越界分离的原因。讨论了TA-9(I)在杂交水稻育种中的应用价值和途径。
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引用次数: 0
[Identification of drought tolerant germplasm and inheritance and QTL mapping of related root traits in soybean (Glycine max (L.) Merr.)]. [大豆(Glycine max (L.) Merr.)耐旱种质的鉴定及相关根系性状的遗传和 QTL 绘图]。
Ying Liu, Jun-Yi Gai, Hui-Neng Lü, Yong-Jun Wang, Shou-Yi Chen

Fifty nine accessions of soybean (Glycine max (L.) Merr.) selected from 301 ones in Huang-Huai-Hai and Middle-Lower Changjiang Valleys were tested in two years for their tolerance to drought by using the mean membership index value averaged over those of plant height,leave number,dry root weight and dry stem and leaf weight. Four most tolerant accessions (Rank 1) and two most sensitive ones (Rank 5) were identified. There existed very significant correlations between drought tolerance and relative values of dry root weight,total root length, and root volume (per plant dry weight basis), respectively,which could be used as root indicators of drought tolerance. The RIL population derived from Kefeng 1 x Nannong 1138-2 was used to analyze the inheritance of the three related root traits by using the segregation analysis of quantitative traits under the major gene plus polygene mixed inheritance model. The results showed that between the two parents (Rank 1 x Rank 4), the relative values of dry root weight,total root length and root volume were respectively controlled by two major genes (linked together for the latter two traits, recombination value being 4.30% and 1.93%, respectively) plus polygenes with their major gene heritability values of 62.26%-91.81% and polygene heritability values 2.99%-24.75%, indicating that the major genes,especially the one with larger effect,accounted for a major part of the genetic variation between the two parents. It was identified that five, three, and five QTLs located on N6-C2, N8-D1b + W, N11-E, and N18-K linkage groups for relative dry root weight, total root length and root volume, respectively. Each of the traits appeared to have one locus (Dw1, R/1, and Rv1) with relatively large effect in comparison with their other loci, and those major ones were located near the same site of the same linkage group N6-C2. The results of segregation analysis and QTL mapping appeared pretty consistent with each other, which could be used as a demonstration of each other.

从黄淮海流域和长江中下游流域的 301 个大豆(Glycine max (L.) Merr.)品系中选出 59 个品系,采用平均株高、叶片数、干根重和干茎叶重的成员指数值,分两年对其耐旱性进行了测试。结果发现了四个最耐旱的品种(排名 1)和两个最敏感的品种(排名 5)。耐旱性分别与干根重量、根总长度和根体积(单株干重)的相对值之间存在非常显著的相关性,可作为耐旱性的根指标。利用克丰1号×南农1138-2的RIL群体,采用主基因加多基因混合遗传模式下的数量性状分离分析,分析了三个相关根系性状的遗传情况。结果表明,在两个亲本(Rank 1 x Rank 4)之间,干根重、总根长和根体积的相对值分别受两个主基因(后两个性状的主基因连接在一起,重组值分别为 4.30% 和 1.主基因遗传力值为 62.26%-91.81% ,多基因遗传力值为 2.99%-24.75% ,表明主基因,尤其是效应较大的主基因占了双亲间遗传变异的大部分。结果表明,相对干根重、总根长和根量分别有 5 个、3 个和 5 个 QTL 位于 N6-C2、N8-D1b + W、N11-E 和 N18-K 连接组上。每个性状似乎都有一个位点(Dw1、R/1 和 Rv1)与其他位点相比具有相对较大的效应,而且这些主要位点位于同一连锁组 N6-C2 的同一位点附近。分离分析结果与 QTL 图谱结果相当一致,可以相互印证。
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引用次数: 0
FISH analysis of pachytene chromosome and DNA fiber of telomere sequence in rice (Oryza sativa L. indica). 水稻粗线染色体和端粒序列DNA纤维的FISH分析。
Zong-Yun Li, Rui Qin, Wei-Wei Jin, Zhi-Yong Xiong, Yun-Chun Song

Telomere sequences were analyzed by using pachytene chromosome and extended DNA fiber FISH in rice (Oryza, sativa ssp. indica cv. -Guangluai No.4). Pachytene FISH results showed that most of chromosome ends possess the telomere tandem repeats, but the signals on different chromosomes were not the same in intensity. Fiber FISH results indicated that the longest string of beads was 6.55 microm, while the shortest one was 1.82 microm long,which were equal to 16.44 and 4.56 kb correspondingly based on a stretching factor of 2.51 kb/microm. The average value of signal length was 3.62 +/- 1.32 microm, i.e. 9.09 +/- 3.31 kb. It could be estimated that the longest and the shortest as well as the average value were equal to 2,349,651 and 1298 +/- 473 of copy number respectively.

利用粗线染色体和延伸DNA纤维FISH对水稻端粒序列进行了分析。籼稻简历。-Guangluai第四)。Pachytene FISH结果显示,大多数染色体末端具有端粒串联重复序列,但不同染色体上的信号强度不相同。Fiber FISH结果表明,最长的珠串长度为6.55微米,最短的珠串长度为1.82微米,按2.51 kb/微米的拉伸系数计算,珠串长度分别为16.44和4.56 kb。信号长度平均值为3.62 +/- 1.32微米,即9.09 +/- 3.31 kb。可以估计,最长和最短的拷贝数和平均值分别为2,349,651和1298 +/- 473。
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引用次数: 0
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Yi chuan xue bao = Acta genetica Sinica
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