clone molecular dna 中文意思是什麼
clone molecular dna
解釋
克隆分子dna-
In this study, we designed a pair of primers based on the sequence of the upstream and downstream of chicken il - 2 gene. about 600 bp chicken il - 2 cdna fragment was cloned from cona - stimulated chicken splenocytes by reverse transcription - polymerase chain reaction ( rt - pcr ) and was subcloned into puc18 vector. recombinant clone was demonstrated by restriction enzyme digestion and dna sequencing. next, we construct recombinant plasmid pproex ? t - il - 2. the cdna of chicken il - 2 gene was subcloned into bamh i / hind iii sites of vector. the recombinant plasmid pproex ? t - il - 2 was transformed into e. coli dh5a and the bacteria was induced with iptg. it was demonstrated by sds - page and western blot that a 18kda protein which was equal to chicken il - 2 protein in molecular weight was expressed in e. coli dh5a. the expression level was up to 30 % of the total bacterial proteins. the purified protein was used to prepare the antibody against chicken il - 2 protein
經酶切鑒定及dna序列測定，該基因為雞il - 2基因，其序列與sundick等報道的完全一致。在此基礎上，我們把雞il - 2基因亞克隆到大腸桿菌原核表達載體pproex ~ （ tm ） ht中，構建重組表達質粒並進行確證性序列測定，重組質粒測序結果表明將編碼雞il - 2成熟蛋白的基因正確地插入到原核表達載體pproex ~ （ tm ） ht的目的位點。重組質粒轉化受體菌dh5後用iptg於37進行誘導培養， sds - page和westernblot分析顯示，表達的雞il - 2融合蛋白分子量約為18kda ，表達的融合蛋白經薄層掃描發現目的蛋白表達量約占菌體蛋白的30 。 -
And the two pah ' s of pcr primers that bind to the adapter and the sequence of f fragment close by tn5 respectively were also designed. the genomic dna of b8 was isolated, digested with bamh i, and ligated to the adapter. using the two pairs of the primers, two rounds of pcr were performed hi turn and a fragment of 239bp was amplified successfully. lt was proved by cloning and sequencing that 18bp of the fragment is the sequence opposite to f fragment on the left of tn5 insertion site in b8f, the other is part of the 728 bp of f fragment. this result makes it possible to continue to carry out chromosome walking, to clone and sequence the whole genes of b fragment and f fragment, and to reveal the antagonistic molecular mechanism of b8
試驗研究設計併合成了由40和44個堿基的寡聚脫氧核苷酸組成的染色體爬行接頭，在接頭序列和測定的f片段近tn5的序列上，設計了2對染色體爬行用的pcr引物，從b8菌株中提取基因組dna ， bamhi酶切，與染色體爬行接頭連接，依次用2對引物進行pcr ，擴增出239bp產物，經克隆、測序，發現其中18bp為擴增的相應于f片段在b8f菌株tn5插入位點對面的序列，其餘則為f片段728bp序列的一部分，為進一步進行染色體爬行，克隆和測定整個b和f基因，揭示陽菌株的拮抗分子機制提供了技術資料貯備。
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