顯核菌目 的英文怎麼說
中文拼音 [xiǎnhéjūnmù]
顯核菌目
英文
caryophanales-
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 。On the bases of designing a primer pair, we obtained the coding domain sequence of rbp by pcr from cdna library. then the gene was cloned into pgem - t vector, the dna sequencing showed that the cloned gene was in agreement with the reported sequence. then the tageted gene of rbp was further subcloned into a procaryotic expression vector pbv220 and constructed recombinant plasmids was named pbv220 - rbp. in order to expresse rbp in procaryotic cell efficiently, the recombinant plasmids was introduced into e. colidhs a straints. expression of rbp was induced by temperature induction
本實驗在合成該蛋白基因上下游引物的基礎上，利用pcr技術，從人睪丸cdna庫中釣取目的基因，並克隆于pgem - t載體中，經序列測定證明與文獻報道基本一致，再將目的基因從重組克隆質粒中亞克隆于pbv220原核表達載體中，轉化宿主菌，經溫度誘導后，進行sds - page電泳分析，發現在約21kd位置上出現了一條明顯的蛋白帶，與預期相符。
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