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To study the molecular mechanism of Larix somatic embryogenesis, a differentially expressed cDNA library of Larix somatic embryo in the period of maturation was constructed using suppression subtractive hybridization. The cDNA from the cultures at the stage of somatic embryo maturation of embryogenic cell line Y35 of L. leptolepis × L . princi-pis-rupprechtii was used as the tester and the cDNA from its subcultured callus was used as the driver. Eight hundreds ran-domly selected positive clones were sequenced, and 468 UniGenes were obtained finally.

为研究落叶松体细胞胚胎发生的分子机理，文章以日本落叶松×华北落叶松杂种无性系胚性细胞系Y35体细胞胚成熟阶段培养物的cDNA为实验组，继代培养阶段胚性愈伤组织的cDNA为对照组，利用抑制性消减杂交技术(Suppression subtractive hybridization， SSH)构建了体细胞胚成熟阶段的差异表达基因文库。

A total of 13 bands were obtained and 6 of them (A1, A3, A4, A5, A9, A10) were sequenced. The sequences are similar to Leuconostoc mesenteroides (GenBank Access No.AY453065), some uncultured bacteria (AJ318147, AF227834, AJ576427), Ethanologenbacterium (AY434722), Clostridiaceae (AB084627), etc.

共得到13个可辨晰的SSCP条带，对其中的6条带(A1，A3，A4，A5，A9，A10)进行了测序分析，分别同嗜柠檬酸明串球菌(GenBank登录号：AY453065，下同)、未培养细菌(AJ318147，AF227834，AJ576427)、产乙醇杆菌(AY434722)、梭杆菌(AB084627)等相似性较大。

Signal peptide sequence was removed lest it couldnt be recognized by Bacillus subtilis, gene coding for mature peptide was ligated to downstream of sacB signal peptide sequence of pWB980 to form a new ORF and generate pWB980-LipA, gene coding for its chaperone was also amplified and ligated to the downstream of LipA to generate a secretion/expression vector pWB980-LipAB.thechaperone gene was sequenced and analyzed by multiple alignments, resulted showed that there were mutations of nucleotides(1～7) and amino acids (1～2) with the other reported genes.

为了防止枯草芽孢杆菌信号肽酶不能识别脂肪酶的信号肽，切除掉脂肪酶的信号肽编码序列，与枯草芽孢杆菌分泌表达质粒pWB980连接并与质粒上的SacB信号肽构成一个完整的开放式阅读框，获得重组质粒pWB980-LipA，因为该脂肪酶的活性表达需要一个特异性分子伴侣帮助折叠成有活性的构象，将脂肪酶分子伴侣基因串连到脂肪酶基因的下游，获得重组分泌表达载体pWB980-LipAB并且测序分析分子伴侣的基因序列和氨基酸序列并与其它报导的序列进行了比对，发现有1到7个碱基的差异和1到2个氨基酸残基的突变。

Sensory nerve conduction velocity and amplitude of action potential were within the normal range. Sural nerve biopsy was performed in the proband. Berardinelli-Seip congenital lipodystrophy 2 (BSCL2) gene were sequenced in the proband and other 4 patients.

对先证者进行腓肠神经活体组织病理检查，并对先证者和其他4例发病者进行血Berardinelli-Seip先天性脂肪营养不良基因2(BSCL2)基因检查。

402 Bp length fragments of mitochondrial cytochrome b gene from 9 species of genus Lutjanidae in South China Sea were cloned and sequenced.The resulting data were combined with homologous sequences of 9 species in western Atlantic in America and 1 species in Philippines of Lutjanidae downloaded from GenBank to form the analysis matrix.

测定了9种中国南海的笛鲷属鱼类的细胞色素b基因的部分序列，结合来自GenBank中1种分布于菲律宾和9种分布于美国大西洋的笛鲷属鱼类的相应同源序列，用邻接法和最大简约法构建分子系统树。

The PCR technique was used to amplify rDNA-ITS-1 of Lutjanus fulviflamma , then the purified PCR productions were cloned into T-vector and sequenced by M13+/-primers.

以特异性引物扩增了金焰笛鲷的核糖体第一转录间隔区（ITS-1），扩增产物经克隆后测序，测得ITS1长度为566 bp。

The results showed that the optimized content of Mg~(2+) was 5mmol/L; dNTP was 200μmol/L; Primer was 0.5μmol/L; Taq polymerase was lU/50ul in the PCR system. Only one fragment of about 270bp was amplified within the genome of 12 wild species and 23 cultivars of Mallus Mill., which indicated that Ty1-copia-like retrotransposons were archaic components of apple genome. The nested insertion of near and within the reverse transcriptase gene of Ty1-copia-like retrotransposons wasn\'t found in apple genome.2 The reverse transcriptase conservative region of Ty1-copia-like retrotransposons were amplified from Gala apple using the optimized PCR system. The amplification product was isolated, cloned and sequenced. Forty-five clones containing reverse transcriptase conservative region were obtained (accession number: AY849580 -AY849592 and DQ105035-DQ105066). Cluster analysis of these sequences showed a great heterogeneity among RT domains isolated from the same genotype.

从苹果属12个野生种和苹果23个栽培品种中都扩增到了约270bp的目的产物，且所有试材的扩增结果一致，均无目的条带以外的产物扩增，说明Ty1-copia类逆转座子在苹果属植物中广泛存在，且存在历史可能较为久远；从扩增结果看不出该逆转座子的逆转录酶基因或附近区域在苹果基因组内有嵌套行为的发生。2、利用优化了的PCR方法从嘎拉苹果基因组克隆了45条Ty1-copia类逆转座子逆转录酶保守序列(登陆号分别为AY849580-AY849592和DQ105035-DQ105066)，结果表明，同一基因组来源的这些序列存在高度的异质性。

The DNA sequences around the transposon inserted site were also amplified by inverse PCR and sequenced. Multisequence alignments showed that the mutated genes were highly similar to the malonyl coenzyme A transacylaseg gene, lipopeptide synthetase B gene and sensor histidine kinase gene, respectively.

反向PCR克隆转座插入位点周边序列及测序结果表明，转座突变的基因与编码脂肽类化合物合成和调控相关的malonyl coenzyme A transacylase、lipopeptide synthetase B和sensor histidine kinase蛋白基因具有高度同源性。

Two fragments from Malvastrum coromandelianum samples were cloned and sequenced.

对赛葵样本Fz1 DNA-A进行了克隆和序列测定，Fz1 DNA-A全长2741个核苷酸。

Then pMD-P80 is digested by the enzymes Xho I and Apa I, separated and linked to the linear plasmid pEGPF-C1 after digested by Xho I and Apa I. So the recombined expression plasmid pGFP-P80 is achieved. Then it is sequenced, PCR and digested by the restriction enzymes Xho I and Apa I. The results give that pGFP-P80 is achieved successfully and the insert sites, direction and reading frame are all right. It builds the basis for expressing, purifying, gaining p80 protein from mammiferous cells.

将pMD-P80 分别经Xho I 和Apa I 双酶切和回收，然后与经过Xho I 和Apa I 酶解的真核表达载体pEGPF-C1 连接、转化，获得重组质粒，经PCR，Xho I 和Apa I 限制性酶切和序列测定，鉴定为真核表达质粒pEGFP-P80，并且目的基因的插入位置、方向和读码框完全正确，为在哺乳动物细胞中表达并分离纯化p80 蛋白奠定了基础。

Chinese traditional virtue is humility and wariness,the compliment and praise to the others should be declination

中国人的传统美德是谦虚谨慎，对别人的恭维和夸奖应是推辞。如

We bought this house on the never-never.

我们以分期付款的方式买下这座房子。