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Histochemical assays of GUS gene showed those resistant callus and roots from transgenic plants cultured under light were no blue-stained with X-gluc, but callus and roots after 24 h treatment of darknees and then incubated with X-gluc could present blue. Fluorimetric enzyme quantitative analyses indicated GUS expression in transgenic plants was organ specificity.

转基因抗性愈伤和转基因植株根的组织化学GUS活性检测表明，光照培养下的抗性愈伤和转基因植株根不能使X-gluc显色，而暗处理24 h后的抗性愈伤和定植后转基因植株的根能使X-gluc显色。

The PCR products were examined by agarose gel electrophoresis. The target gene fragments were purified by gel extraction kit and ligated to cloning vector pMD18-T. The recombinant vectors were transformed into host strain E. coli K802 by lithium chloride method, screened and identified with PCR and restrictive enzymatic digestion. Their sequences were confirmed by DNA sequencing.(2) sTWEAK1 gene was subcloned into expression vector pProEx HTb and transformed into E. coli BL21. sTWEAK2 gene was subcloned into expression vector pMAL-C2x and transformed into E. coli TB1. The recombinant vectors were screened and identified with PCR and restrictive enzymatic digestion. The recombinant fusion proteins were induced to express with IPTG, detected by coomassie brilliant blue-stained SDS-polyacrylamide gel electrophoresis , and confirmed by Western blot analysis.(3) The sTWEAK1 fusion protein was purified with Ni-NTA Spin Kit.(4) The biological activity was assayed on transformed and tumor cells by microplate photometer after crystal violet or sulfur rodamine B staining.(5) The contents of IL-8 in the supernatant of 1990 cell cultures were determined by ELISA.(6) The morphological changes of the sensitive cells were observed by light and transmission electron microscopies.(7) The cell cycle and apoptotic rate were assayed by flow cytometry in 1990 and M85 cells.(8) The effect of fusion proteins on induction of NF-κB in 1990 and LOVO cells was detected with Dual-Luciferase Reporter Assay system.(9) The TWEAK gene was subcloned into Adeno-X Viral DNA with pShuttle vector and transfected into HEK293 cells by lipofectamine method.

(1)本研究用RT-PCR方法，从人组织细胞总RNA中扩增可溶性TWEAK胞外区（sTWEAK1和sTWEAK2）的cDNA序列及全长编码序列，用琼脂糖凝胶电泳分析PCR产物，胶回收目的基因片段，连接到pMD18-T克隆载体中，转化大肠杆菌K802，PCR和酶切筛选阳性克隆，全自动DNA测序验证序列；(2)sTWEAK1和sTWEAK2分别亚克隆到pProEx HTb和pMAL-C2x表达载体中，分别转化大肠杆菌BL21和TB1，PCR筛选和酶切鉴定，阳性克隆用IPTG诱导表达，表达产物用SDS-PAGE分析和Western blot验证融合蛋白；(3)用NTA-Ni Spin试剂盒初步分离纯化sTWEAK1融合蛋白；(4)用体外培养的肿瘤细胞和正常对表达产物进行活性检测，贴壁细胞用结晶紫染色法，悬浮细胞用磺酰罗丹明B染色法，酶标仪检测OD值；(5)敏感细胞用ELISA法检测细胞培养上清中IL-8的含量；(6)用光镜和电镜观察敏感细胞死亡和细胞凋亡情况；(7)用流式细胞仪分析表达产物对敏感细胞凋亡率和细胞周期的影响；(8)用双荧光素酶报告基因检测法，测定表达产物对敏感细胞NF-κB的影响；(9)用pShuttle穿梭质粒将TWEAK重组到腺病毒载体上，用脂质体转染法转染HEK293细胞，PCR鉴定重组质粒。

The cells treated with hypoxia and TRPC3 false siRNA exhibited such apoptotic morphological features as cell shrinkage, condensation, aggregation of nuclear chromatin and presence of bright blue-stained apoptotic body.

荧光显微镜下观察，常氧对照组胞核均匀蓝染；缺氧处理组和缺氧＋TRPC3假干扰组胞核明显固缩、凝聚，可见凋亡小体；而缺氧＋TRPC3干扰组胞核均匀蓝染。

But what strikes one most about the Strip chapels, with their wishing wells and stained-glass paper windows and their artificial bouvardia, is that so much of their business is by no means a matter of simple convenience, of late-night liaisons between show girls and baby Crosbys.

说到这些沿公路商业区的小教堂，以及里面的许愿井、贴有染色玻璃砂纸的窗户，还有他们的人造新娘捧花，最令人惊奇的事莫过于他们的业务绝不是简单的提供方便，或是歌舞女郎和花花公子们的午夜结合。

But what strikes one most about the Strip chapels, with their wishing wells and stained-glass paper windows and their artificial bouvardia, is that so much of their business is by no means a matter of simple convenience, of late-night liaisons between show girls and baby Crosbys.

但赌城教堂最令人称奇的是，除了许愿井、花色玻璃纸窗户以及它们的假花束之外，它们这一行业的相当一部分绝非只是作为一种权宜之计、只是在深夜为那些表演女郎和小克罗斯比[6]们牵牵红线而已。

But what strikes one most about the Strip chapels, with their wishing wells and stained-glass paper windows and their artificial bouvardia, is that so much of their business is by no means a matter of simple convenience, of late-night liaisons between show girls and baby Crosbys.

拉斯维加斯大道的小教堂有许多许愿井、彩色玻璃纸窗户及寒丁子假花，但最引人瞩目的是，它们业务繁多，但绝不仅仅是为了方便客户而已，也不是歌舞女郎与年轻歌手在深更半夜的暧昧。

But what strikes one most about the Strip chapels, with their wishing wells and stained-glass paper windows and their artificial bouvardia, is that so much of their business is by no means a matter of simple convenience, of late-night liaisons between show girls and baby Crosbys.

但是长街上的教堂最让人印象深刻的地方在于它们大多数的业务绝不只是带来单纯的便利，绝不只是关乎深夜里艳舞女郎与克罗斯比家族13败家子们之间的暧昧纠缠。教堂设有许愿水井，有嵌着彩色玻璃、贴着窗纸的窗户，还有人工培植的寒丁子花14。

But what strikes one most about the Strip chapels, with their wishing wells and stained-glass paper windows and their artificial bouvardia, is that so much of their business is by no means a matter of simple convenience, of late-night liaisons between show girls and baby Crosbys.

许愿池、彩色玻璃纸窗、人造布瓦尔花，拉斯维加斯大道的这些教堂给人印象最深的就是它们所做的那么多生意绝不是为图个方便、为了便于让歌舞女郎和像克罗斯比老弟那样的人在深夜私通。

Tom and Summer try their best to find architectural relatives of the Chicago School in downtown Los Angeles: soot-stained but broad-shouldered and highly decorated mid-rise buildings like the Eastern Columbia, in particular.

Tom和Summer在洛杉矶市区努力寻找那种和芝加哥学派有关的建筑，特别是像Eastern Columbia大楼这样楼顶宽大高度装饰的被烟熏暗的建筑。

And rose even more effemination, do not let the water wash-shell lei, once stained, and perhaps the entire bud will rot away.

而玫瑰则更娇气，千万别让苞蕾沾上水，一旦沾上了，整个花苞说不定都会烂掉。

And Pharaoh spoke to Joseph, saying, Your father and your brothers have come to you.

47：5 法老对约瑟说，你父亲和你弟兄们到你这里来了。

Additionally, the approximate flattening of surface strip using lines linking midpoints on perpendicular lines between geodesic curves and the unconditional extreme value method are discussed.

提出了用测地线方程、曲面上两点间短程线来计算膜结构曲面测地线的方法，同时，采用测地线间垂线的中点连线和用无约束极值法进行空间条状曲面近似展开的分析。