请帮我把这个翻译成中文

Recombinant Heparinase (r-Heparinase). r-Heparinase was expressed using the pET system (21), where expression is driven by bacteriophage T7 polymerase. The host E. coli strain BL21(DE3) contains a chromosomal copy of the T7 polymerase gene under the control of lacUV5. The expression is induced by isopropyl f3-D-thiogalactoside. Two constructs were designed to express heparinase in the pET system. The first construct included the native heparinase leader sequence. The second construct started with a sequence that read Met, Gln22, Gln23, Lys24, Lys2s, Ser26 ....
The Met residue was added before the Gln22 to introduce a start codon. Nde I and BamHI restriction sites were appended onto the 5' ends of the N- and C-terminal primers, respectively. F. heparinum genomic DNA was used as a template for 10 scaled-up PCR cycles (see Amplification of the PCR Product) to generate the modified heparinase I product. The PCR product was isolated as discussed above and concentrated using a Centricon P-100 (Amicon) at 5000 x g for 20 min. The amplification product was treated with Klenow fragment of DNA polymerase I (New England Biolabs) for 15 min and with T4 DNA polymerase (New England
Biolabs) for 10 min. The PCR product was isolated on an agarose gel as described above. The blunt PCR fragment was concentrated using a Centricon P-100 and ligated with 20 ng of Sma I-digested pUC18 (Pharmacia) (19). The ligation mixture was then used to transform DH5a-competent cells (GIBCO/BRL/Life Technologies), as recommended by the manufacturer. Subcloned heparinase PCR fragments were excised from pUC18 by digestion with Nde I and BamHI, gel purified, and then ligated to pET-3a plasmid (predigested at the Nde I and BamHI sites and gel purified) using T4 DNA ligase (New England Biolabs). The ligation mixture then was used to transform DHSa-competent cells. The plasmid containing the heparinase I gene in pET-3a was isolated, purified, and used to transform the host cell BL21(DE3) (Novogen,
Madison, WI).
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提示 heparinase:肝素酶 gel:凝胶 plasmid:质粒
recombinant 重组子 host:宿主
机器翻译的就不用发上来了,浪废资源...

第1个回答  2008-05-21
重组肝素酶(的R -肝素) 。的R -肝素有人使用宠物系统( 21 ) ,其中的表达是由噬菌体T7号聚合酶。东道国大肠杆菌菌株BL21中( DE3 )中包含了染色体复制的T7号聚合酶基因控制下的lacuv5 。表达是诱导异丙基的F3三维-硫代。二,建构旨在表达肝素酶在宠物系统。第一,建设包括本土肝素领导人序列。第二,建设开始与序列读会见, gln22 , gln23 , lys24 , lys2s , ser26 ....
该会见了残留之前添加gln22引入起始密码子。我和无损检测的BamHI限制用地附加上5 '两端的N端和C端引物,分别。楼heparinum基因组DNA被用来作为模板10规模的后续聚合酶链反应周期(见扩增PCR产物) ,以产生改性肝素i产品。 PCR产物分离正如上面所讨论和集中使用centricon个P - 100 ( amicon )在5000 xg 20分钟。扩增产品是治疗klenow片段的DNA聚合酶(新英格兰biolabs )为15分钟,并与T4的DNA聚合酶(新英格兰
biolabs )为10分钟。 PCR产物,是孤立于1琼脂糖凝胶电泳如上文所述。该钝的PCR片段,集中使用centricon个P - 100和结扎20霭仪议员的形状记忆合金的I -消化质粒pUC18 (法玛西亚) ( 19 ) 。结扎混合物,然后利用变换dh5a主管细胞( gibco /肝/生活技术)所建议的那样,制造商。亚克隆肝素的PCR片段,切除从质粒pUC18消化与我和无损检测的BamHI ,凝胶纯化,然后结扎宠物- 3A型质粒(简化在无损检测,我和网站的BamHI和凝胶纯化)使用T4的DNA连接(新英格兰biolabs ) 。结扎的混合物,然后用变换dhsa主管细胞。质粒载有肝素酶I基因在PET - 3A型分离,纯化,并用来改造宿主细胞的表达( DE3 )中( novogen ,
威斯康辛州麦迪逊) 。
第2个回答  2008-05-25
再组合Heparinase (r-Heparinase)。 使用宠物系统(21), r-Heparinase被表达了,其中噬菌体T7聚合酶驾驶表示。 主人大肠埃希氏菌张力BL21 (DE3)包含T7聚合酶基因的一个染色体拷贝受lacUV5的控制。 异丙基f3D THIOGALACTOSIDE导致表示。 二修建被设计用宠物系统表达heparinase。 第一修建包括当地heparinase前导序列。 第二修建开始了以读见面的序列, Gln22, Gln23, Lys24, Lys2s, Ser26….
The遇见的残滓在Gln22之前增加介绍起始密码子。 Nde我和BamHI制约站点被添附了在N-和C终端底漆上的5个'末端,分别。 引起修改过的heparinase我产品, F. heparinum genomic脱氧核糖核酸为10个scaled-up PCR周期被用于,当模板(参见PCR产品的放大作用)。 PCR产品被隔绝了如上所述和集中使用在5000 x g的Centricon P-100 (Amicon) 20 min。 放大作用产品对待了与脱氧核糖核酸聚合酶的Klenow片段我(新英格兰Biolabs) 15的极小和与T4脱氧核糖核酸聚合酶(新英格兰
Biolabs) 10 min。 PCR产品在琼脂糖胶凝体如上所述被隔绝了。 直言的PCR片段被集中了使用Centricon P-100,并且绑扎与20 ng Sma我消化了pUC18 (Pharmacia) (19)。 结扎术混合物然后被用于变换DH5a能干细胞(GIBCO/BRL/Life技术),如建议使用由制造商。 Subcloned heparinase PCR片段从pUC18被切除了由与Nde的消化我,并且BamHI,形成胶冻被净化的,然后被绑扎的宠物3a质粒(预先消化在Nde我和被净化的BamHI站点和胶凝体)使用T4脱氧核糖核酸连接酶(新英格兰Biolabs)。 结扎术混合物然后被用于变换DHSa能干细胞。 包含heparinase的质粒我基因在宠物3a被隔绝,被净化了,并且使用变换寄主细胞BL21 (DE3) (Novogen,
Madison, WI)。
第3个回答  2008-05-21
伱疯了吗?
第4个回答  2008-05-21
你傻啊?