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1.
小麂Sry基因的克隆和测序   总被引:5,自引:0,他引:5  
鲁晓瑄  张悦  单祥年 《遗传》2003,25(3):299-301
应用人的性别决定基因SRY(Sex-determining Region Y gene,SRY)中HMG框内的一对引物,对小麂细胞株的基因组DNA进行PCR扩增,得到雄性小麂细胞的220bp扩增产物,而在雌性小麂细胞中未发现扩增产物。将雄性小麂细胞的220bp扩增产物通过T-A互补法克隆到质粒pGEM-T 载体中,筛选阳性克隆进行DNA测序。测序结果表明小麂Sry基因保守序列与人的SRY基因保守区相同碱基的比值为152/184,达到82.6%。提示小麂Sry基因与人的SRY基因存在着较高的同源性,说明SRY基因在进化过程中高度保守。 Abstract:Using the primers from SRY gene——HMG Box for PCR amplification in genomic DNA of Muntiacus reevesi cell strains,a 220bp fragment was obtained in the male but not in the female.The 220bp fragment was cloned into the pGEM-T vector using T/A clone method.The identified positive clone was sequenced.The result shows that 82.6% nucleotides(152bp/184bp) are homologous between Muntiacus Sry and human SRY gene.It suggests that SRY is highly conserved during evolution.  相似文献   

2.
戴君勇  曹祥荣  石磊  张锡然  徐春茂  胡均 《遗传》2005,27(1):95-100
运用PCR扩增、T-A克隆、测序等技术,获得黑麂,小麂,赤麂和毛冠鹿4种麂亚科动物基因组DNA的钾离子通道部分序列。序列分析表明:麂属动物之间的外显子区域序列差异为0.90%~1.44%,毛冠鹿与麂属动物之间的差异为0.90%~1.26%,可见这一段序列的同源性较高。而内含子部分序列差异在麂属动物之间的差异为0~1.22%,毛冠鹿与麂属动物的差异为1.83%左右。由NJ法和最大简约法(Mp法)构建的进化树表明黑麂与赤麂亲缘关系较近,小麂与他们同为一属但关系较远,而毛冠鹿与它们之间的分化程度已达到属间水平。研究表明基因组的内含子序列能够较真实反映近缘动物之间的关系,是进行分子进化比较分析的较理想标记。 Abstract: In this study, partial fragments of potassium ion channel gene were amplified using the genomic DNA of muntjak, reevesi, crinifrons, and Elaphodus cephalophus. The PCR products were ligated to the plasmid of pMD18-T Vector by the method of direct T-A cloning. The positive clones were identified by colony PCR. The sequences of the recombinant clones were determined using M13-47/RV-M universal primers and aligned by the software CLUSTALW. The nucleotide divergences of exon were 0.90%-1.44% among three species of Muntiacus, 0.90%-1.26% between E. cephalophus and each of Muntiacus deer. In the nucleotide of intron there is 0%-1.22% difference among these muntjac deers, and the divergene reached about 1.83% between E. cephalophus and the three species of Muntiacus. Using the software of MEGA to analyse molecular phylogeny, Phylogenetic trees were constructed with neighbor-joining method and maximum parsimony method. The result showed Muntiacus, crinifrons is most closely related to muntjak, with reevesi as their sister species. E. cephalophus is in the other genus.  相似文献   

3.
应用显微切割技术获得赤麂1号,Y1,Y2染色体,通过DOP-PCR增加模板DNA拷贝数,然后用人的性别决定基因(Sex-tetermininig Region of the Chromosome Y,SRY)中HMG框内设计1对引物,对DOP-PCR产物进行扩增,在雄性赤麂Y2染色体DOP-PCR产物中扩增出与人SRY基因同源的Sry基因片段,克隆,测序,首次在分子水平上证明赤麂Y2染色体是真正的Y染色体,同时对赤麂Syr基因进行了初步定位。  相似文献   

4.
采用针对人SRY基因及X与Y染色体同源序列的两对引物进行多重聚合酶链反应技术检测204例新生儿脐血DNA,男性均显示590bp、 355bp及280bp 3条扩增带,女性只有590bp1条扩增带,性别鉴定准确率为100%。又检测47例性器官异常患,13例社会性别为女性者只出现590bp扩增带,与核型性别结果一致。 34例社会性别为男性者中,有2例SRY基因检测结果阴性,只出现590bp带,核型为46,XX,证明这两例患者社会性别不相符,经病理证实后应诊断为女性假两性畸形。 Abstract:This investigation adapted multiplex PCR technique with two pairs of primer to determine sex according to the human SRY gene and X,Y chromosome analogical sequence.We detected bellybutton blood DNA from 204 newborns.Male revealed three bands:590bp,355bp and 280bp,and females only had one band which was 590bp,the accuracy of sex determination was 100%.47 sexual abnormal patients were tested.The results showed that 13 cases,whose social sex were female,had one 590bp band which consistent with their chromosome type sex.In the rest 34 cases,whose social sex were male,2cases showed,on the contrary,only one 590bp band and their chromosome type were 46,XX.This proved that the two patients did not consistent with their social sex,and pathological analysis showed that they were female pseud-hermaphroditism.  相似文献   

5.
对麂属(Muntiacus)中的3种动物:赤麂(M.muntjak)(2n-6♀,7♂),小麂(M.reevesi)(2n=46),黑麂(M.crinifrons(2n=8♀,9♂)MDR-1基因(multidrug resistance,多药耐药基因)726bp左右的片段进行了序列分析并根据序列信息建立分子系统树,同时探讨了这3种动物的起源,分类地位及进化关系。  相似文献   

6.
7.
睾丸决定因子基因(Testis-determining factor,TDF)位于Y染色体短臂上,它的表达产物诱导睾丸组织的发生。SRY基因(Sex-determining Region of the Y)位于Y染色体的性别决定区内,许多特征显示SRY就是TDF。我们选用与SRY基因相应的引物,用PCR技术对正常人男女各10例的基因组DNA进行扩增。将特异扩增的男性SRY基因片段重组到质粒PUC12中,得到含有中国人SRY基因片段的克隆,命名为PSY-1、PSY-2。用[~(32)p]标记重组质粒中的SRY基因片段作探针,与PCR结果进行Southern杂交,男性样品均显示特异杂交带,女性样品为阴性。用末端终止法测定克隆的SRY基因片段的全部核苷酸序列为299bp,含有SRY基因中高度保守及功能特异性区域的240bp,我们对此进行了讨论。  相似文献   

8.
A rice (Oryza sativa L.) mutant displaying defects in panicle development was identified among transformants in a transgenic mutagenlzed experiment using an antlsense cDNA library prepared from young rice panicles. In the mutant, the average splkelet number was reduced to 59.8 compared with 104.3 in wild-type plants. In addition, the seed-setting rate of the mutant was low (39.3%) owing to abnormal female development. Genetic analysis of T1 and T2 progeny showed that the traits segregated In a 3 (mutant) : 1 (wild type) ratio and the mutation was cosegregated with the transgene. Southern blot and thermal asymmetric interlaced polymerase chain reaction analyses showed that the mutant had a single T-DNA insertion on chromosome 5, where no gene was tagged. Sequencing analysis found that the transgenic antisense cDNA was derived from a gene encoding an F-box protein in chromosome 7 with unidentified function. This and another four homologous genes encoding putative F-box proteins form a gene cluster. These results indicate that the phenotyplc mutations were most likely due to the silencing effect of the expressed transgenic antisense construct on the member(s) of the F-box gene cluster.  相似文献   

9.
性别决定基因(Sex region of Y chromosome, 人类以SRY,小鼠以Sry表示)的研究进展是近几年来人类在性别决定,性别分化研究中获得的最大的突破性成果,该文从SRY(Sry)发现前关于性别决定因子的研究,SRY(Sry)的确定,小鼠Sry的结构研究,小鼠Sry的表达研究及Sry下游基因的确定等5个方面对小鼠Sry的研究进展进行综述,对进一步深入研究Sry下游基因存在的瓶颈问题人了一定的分析,并提出核移植技术可能对研究Sry的调节及其下游基因所需的特殊实验材料展现了新的希望。  相似文献   

10.
为确定一个X染色体显性遗传先天性眼球震颤家系的致病基因与X染色体的连锁关系, 选用X染色体上的DXS1214、DXS1068、DXS993、DXS8035、DXS1047、DXS8033、DXS1192和DXS1232共8个微卫星DNA标记对该家系进行基因扫描与基因分型,并利用LINKAGE等软件包对基因分型结果进行分析,探讨该家系致病基因与X染色体的连锁关系。 两点连锁分析时X染色体短臂4个基因座最大LOD值均小于-1,不支持与该家系致病基因连锁; X染色体长臂4个基因座中最大LOD值达到2,提示存在较大的连锁可能性。该家系的致病基因可初步定位于X染色体长臂,且提示Xq26-Xq28区间附近可能是先天性眼球震颤一个共同的致病基因座,但区间范围仍较大,仍须进一步选择合适的微卫星标记进行精确的定位以缩小候选基因的筛查范围。Abstract: To investigate the relationship between X chromosome and obligatory gene of a pedigree with congenital nystagmus,we used the following markers: DXS1214、DXS1068、DXS993、DXS8035、DXS1047、DXS8033、DXS1192 and DXS1232.Genome screening and genotyping were conducted in this pedigree of congenital nystagmus, and linkage analysis by LINKAGE package was used to determine the potential location. The linkage was not found on the Xp ( All LOD score <-1) but on Xq (the maximum LOD score=2). The related gene of this pedigree was located on the long arm of X chromosome. We demonstrate that Xq26-Xq28 is a common locus for CMN. It bring us closer to the identification of a gene responsible for X-linked CMN.  相似文献   

11.
Huang L  Chi J  Wang J  Nie W  Su W  Yang F 《Genomics》2006,87(5):608-615
The black muntjac (Muntiacus crinifrons, 2n = 8[female symbol]/9[male symbol]) is a critically endangered mammalian species that is confined to a narrow region of southeastern China. Male black muntjacs have an astonishing X1X2Y1Y2Y3 sex chromosome system, unparalleled in eutherian mammals, involving approximately half of the entire genome. A high-resolution comparative map between the black muntjac (M. crinifrons) and the Chinese muntjac (M. reevesi, 2n = 46) has been constructed based on the chromosomal localization of 304 clones from a genomic BAC (bacterial artificial chromosome) library of the Indian muntjac (M. muntjak vaginalis, 2n = 6[female symbol]/7[male symbol]). In addition to validating the chromosomal homologies between M. reevesi and M. crinifrons defined previously by chromosome painting, the comparative BAC map demonstrates that all tandem fusions that have occurred in the karyotypic evolution of M. crinifrons are centromere-telomere fusions. The map also allows for a more detailed reconstruction of the chromosomal rearrangements leading to this unique and complex sex chromosome system. Furthermore, we have identified 46 BAC clones that could be used to study the molecular evolution of the unique sex chromosomes of the male black muntjacs.  相似文献   

12.
对麂属(Muntiacus)中的3种动物;赤麂(M.muntjak)(2n=6♀,7♂),小麂(M.reevesi)(2n=46),黑(M.crinifrons(2♀,9♂)线粒体DNA12SrRNA和细胞色素B783bp左右的片段进行序列分析,并根据序列信息建立分子系统树,同时探讨了这3种动物的起源,分类地位及进化关系。  相似文献   

13.
对麂属(Muntiacus)中的3种动物:赤麂(M.muntjak)(2n=6♀,7♂),小麂(M.reevesi)(2n=46),黑麂(M.cri.nifrons)(2n=8♀,9♂)线粒体DNA 12S rRNA和细胞色素b基因784bp左右的片段和核基因-多药耐药基因(multidrug resistance,MDR1)的728bp左右的片段进行了序列分析,并根据序列信息建立了分子系统树,同时探讨了这3种动物的起源,分类地位及进化关系。  相似文献   

14.
利用已测定的鹿科麂亚科动物小麂、赤麂、黑麂的线粒体全基因组序列,统计它们各自连接在一起的13个蛋白编码基因、22个tRNA基因、2个rRNA基因和1个控制区序列的碱基长度和组成,计算rRNA基因遗传距离,估算分歧时间,比较蛋白编码基因的碱基水平和氨基酸水平上的差异,基于连接在一起的13个氨基酸序列,以羊为外群,通过邻位相连法和最大简约性法构建进化树,探讨小麂、赤麂、黑麂的进化关系。结果表明,小麂是较原始的物种,赤麂和黑麂较为近缘,是从类似小麂的祖先演化而来。  相似文献   

15.
近年来,分子细胞遗传学研究已基本证实了染色体的串联融合(端粒-着丝粒融合)是麂属动物核型演化的主要重排方式。尽管染色体串联融合的分子机制还不清楚,但通过染色体的非同源重组,着丝粒区域的卫星DNA被认为可能介导了染色体的融合。以前的研究发现在赤麂和小麂染色体的大部分假定的串联融合位点处存在着非随机分布的卫星DNA。然而在麂属的其他物种中,这些卫星DNA的组成以及在基因组中的分布情况尚未被研究。本研究从黑麂和费氏麂基因组中成功地克隆了4种卫星DNA(BMC5、BM700、BM1.1k和FM700),并分析了这些卫星克隆的特征以及在小麂、黑麂、贡山麂和费氏麂染色体上的定位情况。结果表明,卫星I和IIDNA(BMC5,BM700和FM700)的信号除了分布在这些麂属动物染色体的着丝粒区域外,也间隔地分布在这些物种的染色体臂上。其研究结果为黑麂、费氏麂和贡山麂的染色体核型也是从一个2n=70的共同祖先核型通过一系列的串联融合进化而来的假说提供了直接的证据。  相似文献   

16.
小麂、黑麂、赤麂精母细胞联会复合体的比较研究   总被引:7,自引:1,他引:6  
本工作以界面铺张——硝酸银染色技术,对小麂(Muntiacus reeuesi)、黑麂(M.crinifrons)和赤麂(M.muntjak)的精母细胞联会复合体(Syna ptonemal complex,SC)进行亚显微结构的比较研究。结果表明: 1.SC的平均相对长度和臂比指数同有丝分裂细胞相应染色体的数值有很好的一致性。根据SC的相对长度和臂比指数绘制了三种麂的SC组型图。雄性黑麂减数分裂前期形成一个复杂的易位多价体,意味着其核型的演化过程涉及两次染色体易位和一次臂间倒位。 2.在减数分裂前期,性染色体的形态和行为同常染色体的有明显差异,如性染色体嗜银性较强,配对延迟等。XY的配对起始于早粗线期,在中粗线期,Y的全长均同X配对;XY-SC开始解体于晚粗线期。 3.在粗线期,X染色体未配对区域出现自身折叠,形成“发夹”状结构。这种“发夹”结构的形成,可能是在性染色体的进化过程中,X染色体通过不对称易位得到的重复片段在减数分裂前期同源配对的一种细胞学表现。  相似文献   

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