Investigation on agronomic characters and quality characters of these new soybean varieties 大豆脂肪氧化酶缺失體的農藝和品質性狀鑒定
With ni2 + - lda - sepharose , the cbd tag was removed . according to the results of sds - page and western blot analysis , the product of outflow was the target protein lt a 27 . the purity of lt a 27 was about 95 % 運用niz +金屬鰲合親和層析柱進一步快速純化去除融合頭,上樣流出經sds一隊ge分析和western印跡分析ii1 :明得到的純化產物為淋巴毒素缺失體蛋白lt凸27 , lt 2 :產物純度可達95 %以上。
Lymphotoxin ( lt ) is a kind of pleiotropic lymphocyte - secreted cytokine which mediates a large variety of inflammatory , immunostimulatory , and antiviral responses . in order to increase the antitumor activity of lymphotoxin and reduce its side effects , the recombinant plasmid pet36b - lt 27 was constructed to express soluble fusion protein cbd - lt 27 . the active form of lt 27 could be collected directly with several simple steps by three kind of components on the expressed fusion protein 本研究通過構建表達n端缺失27個氨基酸的淋巴毒素融合蛋白的重組質粒,在大腸桿菌中實現融合蛋白的可溶及分泌表達,同時利用表達載體上的幾種特殊序列經簡單的分離純化步驟直接獲得大量的有生物活性的淋巴毒素缺失體lt 27 ,為尋找一種高抗腫瘤活性、低臨床毒副作用的生物抗癌藥物進行了有效的探索。
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 采用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 。
The biological activity of purified lt 27 was tested with the assay system , and its biological activity was 2 - 3 107 iu / mg . pro . the cytotoxicity of purified lt 27 was in the same level with rhtnfp and lt international standard . it shows that lt deletion could keep its high cytotoxicity towards tumour cell l929 in vitro after 27 amino acids deleted from its n - terminal 用建立的淋巴毒素生物活性測定方法對上述純化的淋巴毒素缺失體lt 27的生物活性進行檢測,測得其比活為2一3xl口iu / mg . proo純化的淋巴毒素缺失體lt 27的生物活性與rhtnfp和淋巴毒素國標標準品的生物活性大致相當,表明lt經n端缺失27個氨基酸后仍能保持很高的體外腫瘤細胞毒活性。