基于Fisher判别法的一种DNA序列分类方法

针对DNA序列分类的分属问题,提出采用Fisher判别法进行分类。根据氨基酸分子的性质,构建DNA序列对应的特征向量空间,然后对训练集中的A、B两类DNA序列提取特征,建立Fisher判别函数。应用该判别函数对测试集中的182个DNA序列进行分类实验,结果表明该方法具有很好的分类准确度。     

一个新的DNA计算的图表示

DAN计算是新兴的一种智能计算方法,二维平面中建立合适的图表示方法对研究DNA序列有重要作用.通过修改碱基序列的向量表示,给出了一个新的DNA序列的图表示方法,并运用代数方法,证明了该图表示方法的退化度至少可达1/m4+m3-m2-m-1.仿真实验表明,给出的图表示对区分不同基因序列有效.     

基于支持向量机的DNA序列分类系统的设计与实现

针对传统统计方法进行DNA序列分类时要求DNA序列样本的概率分布函数已知，但多数情况下概率分布函数未知这一问题，采用支持向量机这一新的机器学习方法对DNA序列进行分类；以VB和Matlab为主要工具开发了基于支持向量机的DNA序列分类系统。结果表明：该系统能够动态选择DNA训练样本、待测试样本．以及支持向量机模型中的参数，并根据用户的指定条件动态输出计算结果；对于预测一批已知正确分类答案的DNA序列，系统能够自动统计识别率，以观察参数变化对于算法执行结果的影响。支持向量机能够在概率分布函数未知的条件下对DNA序列进行分类。     

烟曲霉植酸酶基因的克隆及在毕赤酵母的高效表达

根据文献报道的烟曲霉植酸酶序列，设计引物以烟曲霉CCTCCAF93044的总DNA为模板进行梯度PCR，扩增出了1条约1．4kh的带，将该片段进行DNA序列测定，其序列与已报道的烟曲霉植酸酶序列完全一致，但只编码完整的成熟植酸酶序列。将该片段克隆到毕赤酵母分泌型表达载体pPIC9K上，获得表达质粒pHBM108。该质粒转化毕赤酵母KM71所得重组子经PCR验证后进行诱导表达，其中一重组子经144h诱导，植酸酶表达量至少为1．25mg／ml，比同类报道的表达量高出60％以上，以植酸钠为底物测得酶活为11．17U／ml。     

中国白梨S基因研究——Ⅱ9个主栽品种S基因型的确定及S29-RNase新基因的分离鉴定

以中国白梨9个品种为实验材料．在分子水平上对其基因组DNA进行了PCR—RFLP系统检测、DNA序列测定及生物信息学分析．鉴定了6个品种的S基因型和3个品种的一个S等位基因．并从天生伏和蜜梨中分离鉴定了一个新的S等位基因,命名为梨Szg-RNase基因．该基因与S13的DNA序列最接近，推导氨基酸序列与S13仅有2个氨基酸序列的差异．     

一个DNA计算的图表示问题

DAN计算是新兴的一种智能计算方法,2维平面中建立合适的图表示方法对研究DNA序列有重要作用。对他人证明的一种图表示方法的退化度进行了纠正,并给出了反例说明其真实退化度。     

植物遗传研究与分子标记技术

分子标记是DNA分子碱基序列变异的直接反映。利用限制性内切酶，酶切生物体DNA后来检测不同遗传位点等位变异(RFLP)，以一个碱基顺序随机排列的寡核苷酸序列为引物，利用对基因组DNA随机扩增来鉴别DNA多态性(fL~PDs)．真核生物基因组中普遍存在的重复序列产生了微卫星(microsatellites)标记技术.而RFLP与RAPDs有机结合形成AFLP技术SNP标记利用基因位点上等位型(alleles)为DNA芯片技术应用于遗传作图提供了基础。对植物种质资源遗传多样性的全面了解，有益于正确制定遗传资源收集和原位保存的策略，有助于鉴定植物遗传多样性中心等，对于拓宽遗传基础的育种策略十分重要分子标记是检测种质资源遗传多样性的有效工具。     

广东株家蚕微孢子虫特异DNA片段的克隆与序列分析

利用碱裂解方法抽提广东株家蚕微孢子虫(Nosema bombycis Naeegeli，N．b．)基因组DNA，通过PCR技术扩增得到了N．b．的一段特异DNA片段。对该目的片段的序列分析发现，该特异DNA片段大小为317bp，与Malone等报道的日本株N.b．特异DNA片段大小一致，但核苷酸序列同源性仅为92．1％。     

牦牛乳球蛋白基因5＇调控区的分离、克隆及序列分析

根据文献设计1对PCR引物，寡聚核苷酸序列上游引物为：5＇-gCg，AAT，TCA，TCC，CAC，gTg，CCT，gC-3’，下游引物：5＇-gAg，AAT，TCC，Tgg，ggA，ggg，ACE；TT-3’。经68℃退火及30轮循环的PCR程序后扩增出长度为1447bp的DNA片段，将该片段从琼脂糖凝胶中回收，克隆到pMD18-T Vector后进行序列测定。序列分析结果表明，牦牛乳球蛋白基因5’调控区与文献报道的奶牛该基因同源性为97．65％，突出的碱基差异为牦牛乳球蛋白基因5’调控区发生了1个位点连续3个碱基的缺失突变和另一位点1个碱基的插入突变。该研究也为开展以牦牛乳球蛋白基因5’调控区为启动子的乳腺生物反应器研究准备了条件。     

亚麻抗锈病近等基因系RAPD特异指纹带的克隆分析

通过随机扩增多态性DNA对6个亚麻抗锈病近等基因系进行分析，获得了2个稳定的特异指纹带，成功地将这2个特异指纹带克隆入质粒载体，并对它们进行了序列测定，获得了2条亚麻抗锈病近等基因系RAPD特异指纹带的DNA序列。     

A ligase-mediated gene detection technique

An assay for the presence of given DNA sequences has been developed, based on the ability of two oligonucleotides to anneal immediately adjacent to each other on a complementary target DNA molecule. The two oligonucleotides are then joined covalently by the action of a DNA ligase, provided that the nucleotides at the junction are correctly base-paired. Thus single nucleotide substitutions can be distinguished. This strategy permits the rapid and standardized identification of single-copy gene sequences in genomic DNA.     

Rous sarcoma virus nucleotide sequences in cellular DNA: measurement by RNA-DNA hybridization

Kinetic analysis of the hybridization of 71S RNA from Prague strain of Rous sarcoma virus with an excess of DNA from virus induced sarcomas indicated the presence of the majority of the viral genome sequences in cellular DNA with a very low average frequency per cell. About one-third of the viral sequences were at least partially complementary to DNA sequences with a higher average frequency on the order of 50 to 100 per cell. Normal chick embryo DNA was distinctly different, but contained sequences at least partially homologous to some fraction of the viral RNA.     

Molecular Genetics of the Bithorax Complex in Drosophila melanogaster

The bithorax complex in Drosophila melanogaster is a cluster of homeotic genes that specify developmental pathways for many of the body segments of the fly. The DNA of the bithorax complex has been isolated, and a region of 195,000 base pairs that covers the left half of the complex is described here. The lesions associated with many of the bithorax complex mutants have been identified, and most are due to DNA rearrangements. Most of the spontaneous mutants have insertions of a particular mobile element named "gypsy." This element affects the functions of sequences removed from the site of insertion. Mutant lesions for a given phenotypic class are distributed over large DNA distances of up to 73,000 base pairs.     

Recognition of thymine adenine.base pairs by guanine in a pyrimidine triple helix motif

Oligonucleotide recognition offers a powerful chemical approach for the sequence-specific binding of double-helical DNA. In the pyrimidine-Hoogsteen model, a binding size of greater than 15 homopurine base pairs affords greater than 30 discrete sequence-specific hydrogen bonds to duplex DNA. Because pyrimidine oligonucleotides limit triple helix formation to homopurine tracts, it is desirable to determine whether oligonucleotides can be used to bind all four base pairs of DNA. A general solution would allow targeting of oligonucleotides (or their analogs) to any given sequence in the human genome. A study of 20 base triplets reveals that the triple helix can be extended from homopurine to mixed sequences. Guanine contained within a pyrimidine oligonucleotide specifically recognizes thymine.adenine base pairs in duplex DNA. Such specificity allows binding at mixed sites in DNA from simian virus 40 and human immunodeficiency virus.     

A brief review of genome editing technology for generating animal models

The recent development of genome editing technologies has given researchers unprecedented power to alter DNA sequences at chosen genomic loci, thereby generating various genetically edited animal models. This mini-review briefly summarizes the development of major genome editing tools, focusing on the application of these tools to generate animal models in multiple species.     

荣昌猪TYP基因克隆及其序列分析

实验根据人、牛的酪氨酸酶(TYR，tyrosinase)基因序列设计了3对引物，扩增出猪的酪氨酸酶基因外显子2，3，4序列，其长度分别为210，135，182bp。对其进行了克隆测序，序列被GenBank收录，登录号分别为：AY236997，AY279998，AY242973。与人、牛、鼠的酪氨酸酶基因序列比对结果显示，TYR基因该区段在人、牛、鼠、猪上极其保守。     

植物特异DNA序列应用研究进展

植物特异DNA序列是指在某些植物属、种、基因组或染色体上单独具有的特异存在的DNA序列。几乎所有的高等生物基因组中都有一些物种或基因组特异(有)的DNA序列。研究这些特定序列,对研究物种间在进化过程中的关系及现有物种间的亲缘关系、特异性状表达等方面有着重要的意义。简述了植物特异DNA序列的种类,总结了利用特异DNA序列作为与某一性状基因连锁的分子标记的应用情况,以及特异DNA序列在鉴定外源染色体的来源、某一染色体组或基因组、某一物种或属植物等方面的应用情况,提出了存在的问题与应用前景。     

Isolation of human transcribed sequences from human-rodent somatic cell hybrids

A method was developed for selectively isolating genes from localized regions of the human genome that are contained in interspecific hybrid cells. Complementary human DNA was prepared from a human-rodent somatic cell hybrid that contained less than 1% human DNA, by using consensus 5' intron splice sequences as primers. These primers would select immature, unspliced messenger RNA (still retaining species-specific repeat sequences) as templates. Screening a derived complementary DNA library for human repeat sequences resulted in the isolation of human clones at the anticipated frequency with characteristics expected of exons of transcribed human genes--single copy sequences that hybridized to discrete bands on Northern (RNA) blots.     

大豆基因组保守序列及综合保守序列扩增多态性分子标记(ICSAP)的设计

采用VBA程序查找了大豆基因组保守序列——在基因组中出现次数较多的碱基序列片段,分析了这类保守序列的基本特性,在此基础上,设计了一种新型分子标记——综合保守序列扩增多态性分子标记.碱基数目越多,保守序列种类数和出现次数越少,从10个碱基到27个碱基,保守序列的GC含量先是逐渐增加,达到一个平台后又逐渐降低.碱基离保守序列3＇端越近,出现A或T的可能性越大,碱基在保守序列中的分布具有非随机性.不同长度的保守序列具有明显的进化关系.筛选出10个碱基的保守序列和长度为8个碱基的填充序列用来设计引物,共得到566对引物组合.该新型分子标记有望在条带多态性、引物一致性等方面超越现有分子标记,同时具有成本低廉等优点.该研究旨在为基因组保守序列查找和分子标记设计方面提供新的思路,分析这些保守序列特性有助于进一步探索分子进化机理.该新型分子标记有可能在大豆种质资源鉴定和基因图位克隆等方面得到应用,同时该分析方法也适用对其他物种.