端粒擴增法 的英文怎麼說
中文拼音 [duānlìkuòzēngfǎ]
端粒擴增法
英文
telomeric repeat amplification protocol-
As well as in eukaryocyte ( hepg2 and cos - 7 ), then detect their antigenity as a basis study and explore of the choice of immunogen for preventive and therapeutic vaccines of hepatitis b. methods : the gene fragments coding 152aa ( si ) and 124aa ( s2 ) of the carboxyl terminus of hbsag were amplified by pcr from plasmid pecob6 with a pair of primers containing different endonuclease sites and were cloned into multiple cloning sites of plasmid pbks ( + )
為乙型肝炎的預防和治療性疫苗免疫原的選擇進行初步的研究和探討。方法：本研究利用聚合酶鏈反應（ pcr ） ，通過設計帶有不同酶切位點的一對引物，從質粒pecob6特異性擴增hbsag蛋白羧基末端152個氨基酸（ s1 ）和124個氨基酸（ s2 ）的基因片段，分別將二者克隆到質粒pbks （ + ）的多克隆位點，篩選重組克隆。Objective : to clone and sequence the cdna encoding metalloproteinase from the venom of agkistrodon acutus from northen mountain area of guangxi province. methods : one step method was used to extract total rna from the venom of agkistrodon acutus found in northern mountain area of guangxi province. different kinds of cdna encoding metalloproteinase were amplified by one step method ( rt - pcr and pcr reactions occurred in the same tube ) using different primers
方法：從桂北五步蛇毒腺中抽提總rna ，利用不同的引物，採用一步法（ rt - pcr和pcr在同一管內進行）擴增出不同的dna條帶，利用平端連接的方法將pcr擴增產物克隆至pgem - teasy載體，轉化大腸桿菌jm109 ，挑選白色菌落提取質粒，用pcr對其進行鑒定，直接利用純化pcr產物或提取陽性菌落質粒進行測序。A human lymphotoxin deletion gene fragment which lacking n - terminal 27 amino acid residues of the protein was pcr amplified using a pair of designed primers and cloned into expression vector pet36b ( + ) to construct a fusion protein with cbd tag, resulting a recombinant plasmid pet36b - lt 27. the recombinant plasmid was transformed into host e. coli bl21 ( de3 ) plyss
採用pcr的方法擴增出人淋巴毒素n端缺失27個氨基酸的缺失體基因片段，將此基因片段克隆至表達載體pet36b （ + ）上，與t7lac啟動子控制下的cbdtag構建成表達融合蛋白的重組質粒pe736b - lt 27 。After electrophorised on 1 % agarose gel, the pcr production was purified with agarose gel dna extraction kit. the segment was ligated with vector pmd18 - t and then was tranformed into the competent cell of dh5 a. a construction mstnd - pmd18t was generated by inserting the sequence of 254bp into pmd18 - t vector and selecting the sense clones. positive clone was identified by three ways : endonuclease digestion, pcr and sequencing. the result showed that the cloned sequence coincides with the designed sequence. this construction was digested with nco i and xho i and ligated the pet28a ( + ) vector digested with the same enzymes using dna ligation kit. the production of ligation reaction was transformed into the competent cell of bl21 ( de3 ). after 12 - 16 hours of culture, several colnes appeared on the plate. some positive clones were selected to extract their plasmid. these plasmids were digested by nco i and xho i and indentified by pcr. a contraction, mstnd - pet28a was generated. the result showed that the cloned sequence coincides with the designed sequence
F _ 1長38bp ， r _ 1長36bp ，其它片段均40bp長， f _ 1和r _ 1片段兩端分別加上限制性內切酶nco和xho的識別位點序列。用成對單鏈片段進行延伸反應，然後用其他單鏈片段作為引物，進行pcr擴增，用dna快速純化回收試劑盒回收所得254bppcr產物，與pmd18 - t載體連接、轉化dh _ 5 。受體菌感受態細胞，利用藍白斑遺傳學篩選法篩選陽性克隆，提取其質粒，採用nco和xho雙酶切鑒定，獲得了254bp的片段；用pmd18 - t載體上的特異引物rv - m和m13 - 47進行pcr鑒定，獲得300bp的片段。2. 4 preparation of yeast telomerase, the activity of telomerase was examined by the method of trap, polyacrylamide gel electrophoresis and silver staining
2 . 4酵母端粒酶的制備，端粒重復擴增法（ trap ） 、聚丙烯酞胺凝膠電泳及硝酸銀染色法檢測端粒酶活性。That is to add a special fluorescence - based dna internal standard in the telomerase elongated ts primers, then do pcr amplification after a step of refinement ( hydroxybenzene / chloroform extracting, and deposited by ethanol ). sequencing analysis of pcr product was done on 310 gene scan analysis ?. 1. 2 dna sequencer to determine telomerase activity. notably, this method eliminated the restraining factors of taq dna polymerase, making it possible to erase the sample differences met in pcr and eradicate the annoying phenomena of pseudo negative results
在kim等開發的端粒重復擴增分析法（ trap ）的基礎上進行改進，即通過對端粒酶延伸ts寡核昔酸反應產物的精製，消除了pcr擴增中抑制taq酶活性的因素，從而減少了樣品之間pcr擴增上的差異和假陰性現象的發生，提高了判斷樣品端粒酶陰、陽性的準確率和定量的準確性。分享友人