In this paper, first strand cdna of 3abc gene was synthesized using template rna extracted from cells infected with fmdv. the complete 3abc gene about isoobp was amplified by pcr and ligated into pgem - t easy vector. after transforming e. coli dh5 a, ampicillin resistant colonies were isolated and plasmid dna was prepared and analyzed by restriction analysis and pcr. presence of the full length 3abc gene was verified by nucleotide sequence analysis and the plasmid containing the expected sequence was named as pgem - 3abc. comparing the aquired sequence of 3abc with that of reference strains, the homology is more than 99 percent. the pgem - 3abc was digested with sal i and bgl ii and ligated into xho i and bgl ii - digested expression vector ptriex - 4 neo. lt was identified by restriction analysis and pcr and sequencing that this fragment had a 17bp deletion hi the nucleotide sequence 708bp of 3abc gene, which happened to form a terminator codon behind 3ab gene, but it contained the complete open reading frame ( orf ) of 3ab gene. positive clones were selected and induced with lmmol / l isopropyl - d - galactoside ( iptg ), bacteria were detected by sds - page and western blotting after properly treated. the results showed that the 3ab gene expressed successfully in e. coli and 33. 5ku fusion protein can be recognized by the positive bovine serum of fmdv. the amount of target protein is over 26 % of the total bacteria protein by gel thin layer scanning analysis
擴增產物連接到pgem - teasy載體中,轉化大腸桿菌dh5菌株,篩選氨芐青霉素抗性菌落,提取質粒經酶切鑒定、 pcr分析以及確證性測序證明,所克隆的1500bp左
右的片段含有完整的3abc基因,與國外參考序列相比,同源性在99以上。將重組質粒pgem - 3abc和表達載體ptriex - 4neo分別用sal和bgl與xho和bgl消化后,亞克隆3abc基因至原核表達載體ptriex - 4neo中,通過酶切鑒定、 pcr擴增以及序列分析,發現克隆到ptriex - 4neo載體上的片段於3abc基因708bp處出現了17bp的缺失,碰巧在3ab基因后形成一終止密碼子,但3ab基因的閱讀框架完整,選出含有3ab基因完整閱讀框架的陽性克隆,用iptg誘導表達,收集菌液進行sds - page電泳、 westernblotting分析,結果表明, 3ab基因在大腸桿菌中成功表達,其表達產物為分子量33 . 5ku的融合蛋白,並能被口蹄疫病毒陽性血
清識別。經薄層掃描分析,表達量占總蛋白量的26以上。
The most notable yellow mountains pines are : yingkesong greeting pine in front of lion stone at jade screen tower, songkesong farewell pineto the right of jade screen tower, putuansong hassock pine in lotus gully, feng huangsong phoenix pine in heavenly sea, qipansong chessboard pine at pingtian stone bridge, jieyinsong usher pine on beginning - to - believe peak, qilinsong unicorn pine, on the way from north sea guesthouse to cool terrace, heihusong black tiger pine, on the way from north sea guesthouse to beginning - to - believe peak, tanhaisong or wusong probe - the - sea or dancer pine by crucian carp s backon celestialcapital and wolongsong recumbent dragon pine below beginning - to - believe peak
最著名的黃山松有:迎客松位於玉屏樓的石獅前面,送客松位於玉屏樓的
右邊,蒲團松位於蓮花溪谷,鳳凰松位於天海,棋盤松位於平田石橋,接迎松位於始信峰,麒麟松位於北海賓館和
清涼臺之間,黑虎松位於北海賓館和始信峰之間,探海松或叫舞松位於天都峰的鯽魚背旁邊這就是黃山的十大名松。
Stop at the kerb and look both ways before crossing ( the road )
先站在路邊看
清左
右再過馬路
The forms are locked about two in the morning.
清早兩點左
右封版。
On the day of surgery, i got there about 1and 1 2 hour before my schedule. i was quite nervous, the nurses were kind enough to calm me and put anesthetic eye drops to my eyes to prepare me for the operation. the surgery took less than 20 minutes and when i got to the recovery room, i can see very clearly already, i even drove the next day
進行激光矯視手術前,需一個星期不配戴隱形眼鏡,做手術時,醫生將我的眼球進行麻醉,而整個人是
清醒的,手術大約花幾十分鐘便完成,張開眼的時候雖不是完全
清晰看到面前的事物,大慨回復了一半左
右的視力,但回家休息過后,第二晚已經可以自己駕車。
If an initation clearly states “ white tie ” or “ formal ” on the lower right, it is required to wear the formal eening clothes for the man and a ball gown and jewelry for the woman
(如果請帖
右下角
清楚地註明「白領結」或「正式」的話,男士應該穿正式晚禮服,而女士則應著晚禮服並佩帶寶石飾品。 )
If an invitation clearly states “ white tie ” or “ formal ” on the lower right, it is required to wear the formal evening clothes for the man and a ball gown and jewelry for the woman
(如果請帖
右下角
清楚地註明「白領結」或「正式」的話,男士應該穿正式晚禮服,而女士則應著晚禮服並佩帶寶石飾品。 )
Avian encephamyelitis ( ae ), which is also called epidemic tremor, characterized by nonsuppurative encephamyelitis, is caused by avian encephalomyelitis virus and has a sign of decreasing egg production. in adult chickens and decreasing hatching rate in fertilized eggs and has 40 % - 60 % of incidence and 25 % of mortality
該病發病率為40 60 ,死亡率為25左
右。成年雞感染后出現產蛋率下降,種蛋出雛率降低。但雛雞感染aev的早期血
清中的igm , igg系統的變化規律尚未見報道。
Players need to understand the game box colors, then passed above and below key to the movement of the box with the color combination, can cross the border
游戲需要玩家認
清方塊的顏色,然後在通過上下左
右鍵的移動將同顏色的方塊組合在一起,即可過關。
After incubated for 5 days, the majority of plagues was about 1mm in diameter. the shapes of plagues were difform and some of them were round. the plagues were no color under low power microscope and the area of the plagues consisted of obtrite dead cell lumps
採用雙層法用ma104細胞進行病毒蝕斑試驗, jl94能形成大多數直徑約1mm左
右的蝕斑,大部分形狀不規則,少部分呈圓形,鏡下可見
清晰的不著色區,此區由碎裂的死細胞團構成。
Following the celebration, one of the quan yin messengers who hosted the event shared the message of peace with attendees, saying that after the september 11 disaster in the united states, he had made this maxim from master his motto : " that mankind may understand each other through more peaceful ways.
下午,大家在森林大殿集合打坐之後,一起慶祝
清海日,主持的一位觀音使者與大家分享和平的訊息。美國911事件之後,師父對世人的提醒:願人類能藉由更和平的方式彼此互相了解,這句話已成為他的座
右銘。
The products can be emulsified in 70 c hot water, and then the waxes are good to penetrate and adhere the cotton fiber and the waxes can be washed easily after sizing process and without influence to the process of dye uniformity
本產品在70 c左右的熱水中乳化,對棉纖維有良好的滲透性和附著力,完成上漿功能后易清洗,不影響后續勻染工藝。