巴西橡胶树TCTP基因结构分析及分子标记开发

翻译控制肿瘤蛋白(translationally controlled tumour protein,TCTP)是生物体中一种普遍存在且高度保守的蛋白.作为一种多功能蛋白,TCTP在各种复杂的生命活动中发挥重要作用.本研究根据已知的HbTCTP cDNA序列设计引物,对PCR扩增产物测序获得HbTCTP基因组序列.比对HbTCTP基因组和cDNA序列发现HbTCTP由4个内含子和5个外显子组成,内含子与外显子剪接处符合GT/AG法则.根据HbTCTP序列及基因结构信息开发简单重复序列(SSR)和内含子长度多态性(ILP)标记,并在10个橡胶树品系中分析SSR和ILP标记多态性.结果表明,SSR和ILP标记在10个橡胶树品系中均出现多态性,多态性分别由SSR重复次数多少和内含子长短引起.     

Intron Polymorphisms： Species-specific and Interspecies-comparable Markers

As a component of genes, introns are much more variable than exons. Therefore, introns are an important source of genetic polymorphisms and can be used as genetic markers. By desingning PCR primers in the two exons flanking an intron, the intron can be specifically amplified. There are mainly two types of intron polymorphisms： intron length polymorphism （ILP, due to InDel or SSR） and SNP. ILP can be conveniently detected by electrophoresis, while SNP can be detected by sequencing, ECOTILLING or other methods.     

A comparative account of the structure of the growth hormone encoding gene and genetic interrelationship in six species of the genus <Emphasis Type="Italic">Labeo</Emphasis>

The growth hormone (GH) gene isolated and cloned from various Labeo species (L. rohita, L. calbasu, L. fimbriatus, L. gonius, L. bata, and L. kontius) is shown to contain a single copy in the haploid genome, with an overall size of ∼2.5 kb. The GH gene in all the Labeo species studied has five exons and four introns of various sizes with the exon/intron boundary sequence of GT/AG. The length variation of the GH gene between the species is found to be due to length variation in the form of several deletions in the third intron. The length of individual exons is the same in all the species with an open reading frame (ORF) of 630 bp (210 amino acids) except in L. rohita, which has a 9 bp deletion in the fourth exon, resulting in a shorter GH of 621 bp (207 amino acids). The similarity in the nucleotide and amino acid sequences between the different Labeo species is greater than 97%, in spite of eight amino acids being altered in the GH protein of Labeo that reside outside the conserved domain sequence required for its function. Nucleotide substitutions are seen in the form of 20 transitions and three transversions in the ORF of the GH gene. Both types of transitions (A–G; T–C) and only one type of transversion (A–C) are detected in the GH gene. Codon preference in GH gene shows a strong preference for G and C in the wobble position of the codons. Genetic interrelationships determined between Labeo and other species of fishes using nucleotide sequence of GH cDNA supports the overall teleost classification of Nelson (Fishes of the World. Wiley, New York, 1984) with separate clades for Ostariophysi, Protacanthopterygii, and Acanthopterygii. Besides, the unweighted pair group method with arithmetic means (UPGMA) analysis clearly distinguishes between the species having five exons and four introns in the GH gene from the species having six exons and five introns in the same gene. The Labeo species analyzed in the present study could be clustered into two groups using the maximum-parsimony method on the intron sequences data of the GH gene.     

小鼠基因内含子中CpG岛和几个转录调控元件分析

内含子在基因转录调控中的作用已多次被实验报道,然而对其参与调控的普遍性还缺乏足够的理论支持。本研究利用计算分析方法,对小鼠基因内含子中的CpG岛（CpGisland）、TATA框（TATAbox）、CAAT框（CAATbox）以及GC框（GCbox）等元件的出现频率进行分析。结果发现,分别有56.01%、57.16%、65.88%和41.86%的第一内含子具有CpG岛、TATA框、CAAT框以及GC框,而它们在其它内含子中的平均出现频率则分别为14.07%、45.24%、49.91%和13.19%。即使考虑到不同位置的内含子,这些元件在第一内含子中的出现频率也显著高于它们在其它任何位置内含子中的出现频率。由于CpG岛、TATA框、CAAT框以及GC框均与基因的转录调控有关,据此推测小鼠第一内含子在基因转录调控过程中具有潜在的重要性。本研究结果为内含子参与转录调控提供了更多的理论依据。     

黄瓜CSHSP70基因内含子结构分析

 【目的】探明黄瓜CSHSP70基因的内含子数目及其结构特点。【方法】根据黄瓜CSHSP70基因cDNA序列设计引物，应用PCR技术从gDNA中克隆CSHSP70基因，采用RT-PCR技术验证内含子的存在。【结果】经测序与拼接，得到长度为4 056 bp的黄瓜CSHSP70基因，它包含7个内含子。经分析发现这些内含子富含A•T，具有类似启动子和HSE核心结构的序列存在。对黄瓜、甜瓜和丝瓜CSHSP70基因部分序列的比较分析表明，该基因在这3个葫芦科作物中具有很强的保守性。【结论】内含子存在的特殊结构表明其可能对该基因表达具有调控作用。     

The Sequence Variations of Intron-3 of the α-Amylase Gene in Adzuki Bean

This study describes variation of intron-3 of a-amylase gene from 156 breeds of adzuki beansusing SSCP(single-strand conformation polymorphism)analysis. Based on a-amylase gene structure and se-quence, A pair of PCR primers, F (CCTACATTCTAACACACCCT) and R (GCATATTGTGCCAGTACAAT)were designed to amplify intron-3 fragments of a-amylase gene. 14 variant types were detected, including 13,9, 10, 4 variant types in the wild, weed, locally cultivated and modern brought-up adzuki beans respectively,9, 8, 7 variant types of the wild adzuki beans from Japan, China and Korea respectively, and some other va-riant types in the local adzuki beans from China and Bhutan. 60 % of subjects of cultivated races were found tobe EE type in the experiment. In addition, sequence analysis of intron-3 of α-amylase gene from 8 varianttypes reveals the evolution process of various variant types in adzuki beans.     

福州宦溪野生蕉（Musa spp.，AB group）2个PSAG成员的克隆及生物信息学分析

植物光系统Ⅰ反应中心亚基V (photosystem Ⅰ reaction center subunit V,简称PSAG或PS Ⅰ-G)是光合系统Ⅰ的主要组件.具有维持PSⅠ复合体稳定性的重要作用,并与抗盐密切相关.本研究以福州宦溪野生蕉(Musa spp.AB group)叶片为材料,采用同源克隆的方法,首次分离到PSAG基因的2个成员:PSAG1、PSAG2(GenBank登录号分别为JX317082、JX317083),分别为800、827 bp,分别编码150、160个氨基酸;PSAG1、PSAG2的基因组序列分析表明2个成员均没有内含子.生物信息学分析表明:PSAG1、PSAG2具有PSⅠ的X亚基超家族(photosystem Ⅰ reaction center subunitX psaK)保守结构域,是不具有信号肽的跨膜蛋白,具有亲水性:PSAG1、PSAG2均有4个位点发生磷酸化.宦溪野生蕉PSAG在进化过程中形成了特殊的结构特征,即PSAG1和PSAG2没有内含子,并且在不同物种间保守区具有高度的一致性,为保持PSAG功能的稳定性提供了重要保证.     

茶树PCP基因内含子的克隆与序列分析

利用GenBank中公布的茶树花粉壁蛋白基因（PCP）序列设计引物,采用PCR法扩增并测定茶树花粉壁蛋白基因内含子序列。在该基因中发现一个内含子,共207bp,A＋T碱基占67．63％,G＋C碱基占32．37％。经分析发现该内含子具有类似启动子的序列存在,可能对茶树花粉壁蛋白的特异表达有调控作用。     

大豆MYB转录因子基因GmMYBJ6和GmMYBJ7的克隆及表达分析

MYB转录因子以保守的DNA结合域为共同特征, 是植物最大的转录因子家族之一, 广泛参与植物代谢调控。本文根据植物中MYB转录因子的DNA保守结构域, 通过RT-PCR和RACE的方法从大豆品种吉林3号的叶片中分离得到了2个MYB基因GmMYBJ6和GmMYBJ7, 并对其结构特征和表达特性进行了分析鉴定。基因组DNA序列分析表明, 两个基因都含有两个内含子; 酵母系统检测表明, 两个基因均具有转录激活活性; β-半乳糖甘酶活性分析表明, 内含子的存在使两个基因的转录激活活性明显降低; 实时荧光定量PCR检测结果显示, GmMYBJ6在大豆中的表达受ABA、GA3和NAA的诱导, 而GmMYBJ7的表达则受ABA和NAA的诱导; 半定量RT-PCR分析表明, 两个基因在大豆的根、茎、叶、花和未成熟种子中均有表达。GmMYBJ6和GmMYBJ7为典型的R2R3-MYB转录因子新基因; 他们在大豆中的表达可能与ABA和NAA的信号转导途径有关。