三轮

Chicken IL-15(ChIL-15), Which was discovered in 2001, plays a main role in activating NK cells and CD8+ memory T cells, and may therefore have potential as a vaccine adjuvant.According to published ChIL-15 gene sequence, a pair of primers were designed and synthesized to clone ChIL-15 cDNA from ConA-activated chicken splenocytes by RT-PCR. Recombinant plasmid pUC19ChIL-15 carrying the ChIL-15 gene was constructed. The gene encoding mature ChIL-15 (mChIL-15) was amplified from pUC19ChIL-15 by PCR and cloned into pMD 18-T vector. After digested with Kpn I /Pst I , the mChIL-15 gene was subcloned into prokaryotic expression vector pPRoEX橦Ta and transformed into E.coli DH5a .The transformant was induced by IPTG, and the recombinant ChIL-15(rChIL-15) was expressed as a fusion protein with a histidine hexamer tag at the N-terminal end of the protein. Following expression, the protein was purified by ProBond?Nickel-Chelating Resin. The uninduced and induced protein lysates and the purified protein were separated by SDS-PAGE and transferred onto nitrocellulose membrane to identify rChIL-15 by Western blot. The rChIL-15 was used to immune the guinea pig to obtain rChIL-15 polyclonal antibody.

参考已发表的ChIL-15基因序列，设计合成引物，应用RT-PCR技术从刀豆球蛋白A活化的白来航鸡脾淋巴细胞中克隆ChIL-15 cDNA，将其与SmalⅠ酶切处理的pUC19载体连接，构建重组质粒pUC19ChIL-15；用PCR技术从pUC19ChIL-15中扩增出去信号肽的成熟ChIL-15(mChIL-15)基因，与pMD 18-T载体相连构建重组质粒pMDChIL-15；然后用KpnⅠ／PstⅠ双酶切下mChIL-15基因片段，定向亚克隆到经同样酶切处理的带有6个组氨酸标签的表达载体pP_RoEX~HTa中，构建重组质粒pP_RoChIL-15，对其测序确认读框正确后，转入大肠杆菌DH5α中诱导表达并进行纯化，用重组ChIL-15(rChIL-15)免疫豚鼠，制备多克隆抗体；再用HindⅢ／PstⅠ从质粒pP_RoChIL-15上切下mChIL-15基因，定向亚克隆到经同样酶切处理的杆状病毒转移载体pMelBacB中，经酶切、PCR和序列测定鉴定后，与杆状病毒线形化DNABac-N-Blue~(TM DNA共转染Sf9昆虫细胞，经三轮蚀斑纯化，构建重组杆状病毒rBac-ChIL-15，用该重组病毒感染处于对数生长期的Sf9细胞，按不同的时间收取样品，离心后对其上清和沉淀进行SDS-PAGE和Western blot分析。

In qualifying it was all Cavalieri, who went on to TQ all three rounds of Truck Modified and 4wd Modified.

在符合资格的，它是所有卡瓦列里，谁接着就tq所有三轮卡车改装越野车和改装。

Mind you, it didn't seem as much when the draw for the 3rd round pitted Chelsea against current English and European champions Liverpool.

但是要提醒你的是，在第三轮中当时英格兰和欧冠双料冠军利物浦和蓝军的遭遇并不是什么了不起的比赛。

After three rounds of diamonds and five rounds of spades, only a five-card ending is possible - you are forced to give up one of dummy's extended menaces.

三轮方块和五轮黑桃后，只有五张牌的残局形式的可能——你被迫放弃明手的延伸威胁之一。

This has been the 3rd round of venture capital investment that this enterprise acquires.

这已经是该企业获得的第三轮风险投资。

BE , which has chauffeured Hollywood starlets to the Oscars in its 3-wheelers, recently hooked up with netLoan Funding to help make the purchase of an EV more like a conventional car, with monthly payments.

的三轮汽车去参加好莱坞的奥斯卡颁奖典礼，最近，该公司推出了netLoan贷款计划，可以像买普通车一样通过每月付款来买该公司的汽车。

3Rd stage: one of the two concertos with orchestra from group 6, chosen by the jury.

第三轮：评委指定第六组中两首协奏曲中的一首，管弦乐团伴奏

Seventy-six grants of $100,000 each are being disbursed in this, the third round of funding.

在第三轮的资助活动中，每十万美元中有76美元的专款被拨付给。

In Chinese market, sales volume of Foton combine harvester has been in the top place in the same field for 8 consecutive years and its percentage of market share excessed 70%; the market share of Foton big and medium horse power tractors and Five-Star brand tri-wheel motors have been the top place for 5 consecutive years.

在中国市场，福田联合收割机销量连续八年居行业首位，市场占有率超过70%；福田大中型拖拉机和福田五星三轮摩托车连续5年市场占有率行业第一；福田旋挖钻机、履带式起重机、装载机、挖掘机等业务市场影响力不断提升，成为行业强势竞争品牌。

In Madrid, the party mood fizzled out in seconds, when they lost out in the third round.

在马德里，当他们在第三轮中被淘汰时，狂欢的情绪在瞬间消失殆尽。

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％。