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The RT-PCR product was inserted into pTG19-T vector and transformed into E. coli successfully. By blastn, the sequence results of Kunming mus musculus were in complete accordance with the conservative sequence of Genbank NR_003278 (791bp-1153bp). By Blastn in NCBI, the sequence with little difference among animals was confirmed to be conservative. After Blastn, fourteen complete CDS coding for different animals were chosen. According to VECTOR NIT 9.0 software, the similarities between Kunming mus musculus and bos taurus, homo sapiens, erinaceus europaeus, cricetulus griseus, sus scrofa, dasypus novemcinctus, rattus norvegicus, rabbit, equus caballus, macaca fascicularis, didelphis virginiana, monodelphis domestica and vombatus ursinus was 67%, 100%, 100%, 36%, 100%, 100%, 67%, 100%, 100%, 92%, 99%, 99% and 99%. In the phylogenetic tress constructed with the forteen 18S rRNA by Treeview, the Kunming mus musculus clustered with cricetulus griseus, sus scrofa and rabbit, which was nearer to cricetulus griseus and was most far away from macaca fascicularis.(3) After sencodary structure analyses of 18S rRNA of mus musculus, an oligonucleotide fragment for RNAi was designed and synthesized, which was transformed into plasmid, and restriction enzyme analyses and sequencing results should the expression plasmid pGPH1/ GFP/Neo-mouse-sh 18S rRNA were constructed for RNAi successfully.

结果①通过RT-PCR检测显示18S rRNA基因在小鼠卵巢组织和单个GV期、MⅠ期卵母细胞中均有表达，且在未成熟卵母细胞中，MⅠ期的表达明显强于GV期的表达；②RT-PCR产物克隆测序结果显示：昆明小鼠18S rRNA基因保守区序列与基因库序列[NR_003278保守区部分(791bp～1153bp)]完全一致；Blastn比对结果发现：在不同物种中差异较小，选出14种生物18S rRNA全序列经VECTOR NIT 9.0软件分析，提示昆明小鼠18SrRNA与牛、人类、刺猬、中国仓鼠、猪、犰狳、褐鼠、兔子、马、食蟹猴、负鼠、短尾猊、袋熊的18S rRNA的相似率依次为67%，100%，100%，36%，100%，100%，67%，100%，100%，92%，99%，99%，99%；Clustal 1.81和Treeview构建出的分子进化树表明：在上述14种生物中昆明小鼠与中国仓鼠进化关系最近，与兔子、猪聚成一簇，与食蟹猴进化关系最远；③根据18S rRNA二级结构设计并合成RNA干扰寡核苷酸片段，重组质粒经过限制性内切酶及测序表明成功构建了pGPH1/GFP/Neo-mouse-sh 18S rRNA干扰表达质粒。

2Ai. In addition, cell extracts from E. coli JM109(DE3) cells harbouring the recombinant plasmid pAM28 were able to decarboxylate the histidine present in the reaction to histamine, whereas extracts prepared from control cells containing the vector plasmid alone did not.

此外，从大肠杆菌细胞提取物大肠杆菌JM109中（DE3）中细胞重组质粒pAM28窝藏能够decarboxylate在本组氨酸为组胺反应，从控制细胞制备含有载体质粒提取物，而仅仅没有。

The plasmid DNA of Streptomyces ahygroscopicus was extracted by soda split. The plasmid DNA was partial hydrolyzed by Sau3AI, and 3.5~9 kb DNA fragment was wllected by dialyze bag.

采用碱裂解法从不吸水链霉菌提取染色体DNA，用Sau3AI对染色体DNA进行部分水解，利用透析袋法回收3.5~9kb之间的DNA片段。

The plasmid DNA bands were detected clearly by agarose-EtBr gel electrophore sis.Two forms of plasmid DNA molecules,namely covalently closed circular DNA mol e-cules and open circular DNA molecules,were observed under electron microscope.

用琼脂糖-溴化乙锭凝胶电泳分析在两株菌的DNA中鉴别出明显的质粒带，用电镜方法在两株菌的DNA制剂中均观察到共价闭合超螺旋和开放型环状两种构型的DNA分子。

Methods The plasmid pCEP4P53 expressing human p53 protein and the plasmid pCEP4ASCMH expressing human antisense c - myc gene in eukaryocyte were co - transfected to human bladder cancer cell T24 strain using FuGENE6 as a media, and the results were analyzed by MTT method, cell colony - forming test in soft agar medium and agarose gel electrophore...

将可在真核细胞表达P53蛋白的质粒pCEP4P53和表达反义c-myc基因的质粒pGEP4ASCMHC以染试剂Fu-GENE6为介导，同时转染到入膀胱癌细胞T24中，经MTT检测、软琼脂集落生长、琼脂糖电泳等方法分析实验结果。

METHODS: To construct recombinant expression plasmid subtype H1 of SIV HA gene of A/Swine/Guangdong/LM/2004(H1N1).BALB/c mice of 6-8 weeks old were immunized endemically with the recombinant plasmid.The splenocytes from the immunized mice were fused with Sp2/0 myeloma cells after the last immunization.

构建 H1 亚型猪流感病毒A/Swine/Guangdong/LM/2004(H1N1)HA基因表达质粒，基因免疫6～8周龄雌性BALB/c小鼠，加强免疫后取其脾细胞与骨髓瘤细胞 Sp2/0进行融合。

Suppression subtractive hybridization cDNA plasmid libraries were constructed between gastrula embryos and tail bud embryos in gynogenetic gibel carp.739 and 816 PCR positive clones were respectively selected to perform dot blot,and 72 dot blot positive clones and 98 dot blot positive clones were obtained from the SSH plasmid libraries specific for gastrula embryos and tail bud embryos.

构建了雌核发育银鲫原肠期胚胎和尾芽期胚胎间的抑制性差减杂交cDNA质粒文库。对原肠期739个和尾芽期816个PCR阳性克隆进行斑点杂交，得到72个原肠期和98个尾芽期斑点杂交阳性克隆。

Results (1)Multiple copies of pMCLacI/Neo plasmid DNA were intactly integrated in the genomic DNAs of the cell clone.(2) One of lac Ⅰ target genes in the integrated plasmid could be transcribed in the NIH3T3 cells while the other could not.

因此，在这些突变婴儿检测系统中，通过一些不表达的中性靶基因所婴儿检测到的突变规律肿瘤是否代表体内一些重要的功能基因的真实情况，目前尚不清楚［1-3］。

Results (1)Multiple copies of pMCLacI/Neo plasmid DNA were intactly integrated in the genomic DNAs of the cell clone.(2) One of lac Ⅰ target genes in the integrated plasmid could be transcribed in the NIH3T3 cells while the other could not.

因此，在这些突变检测系统中，通过一些不表达的中性靶基因所检测到的突变规律是否代表体内一些重要的功能基因的真实情况，目前尚不清楚［1-3］。

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。

Breath, muscle contraction of the buttocks; arch body, as far as possible to hold his head, right leg straight towards the ceiling (peg-leg knee in order to avoid muscle tension).

呼气，收缩臀部肌肉；拱起身体，尽量抬起头来，右腿伸直朝向天花板（膝微屈，以避免肌肉紧张）。

The cost of moving grain food products was unchanged from May, but year over year are up 8%.

粮食产品的运输费用与5月份相比没有变化，但却比去年同期高8％。