JIN Xiao-xia,YU Hai-feng,YU Li-jie.Study on Agrobacterium mediated Inter leukin4 Gene Transferred into Cabbage[J].Northern Horticulture,2013,37(15):113-116.
农杆菌介导的人白细胞介素-4基因转化甘蓝的研究
- Title:
- Study on Agrobacterium mediated Inter leukin4 Gene Transferred into Cabbage
- 文章编号:
- 1001-0009（2013）15-0113-04
- 关键词:
- 甘蓝; 白细胞介素4(IL-4); 遗传转化
- Keywords:
- ; Brassica oleracea L.; IL-4; genetic transformation
- 分类号:
- S 635
- 文献标志码:
- A
- 摘要:
- 以结球甘蓝‘92-1012-3-11’自交系种子转白细胞介素4(IL-4)基因T-0代材料为试材，研究了通过农杆菌介导法将人IL-4基因导入甘蓝后代的遗传转化情况。结表明：共得到抗草铵磷（PPT）抗性再生植株396株，聚合酶链式反应（PCR）阳性植株137株，其中带柄子叶转化率为8.1%，下胚轴分化率为58%。通过对转基因阳性株T0代的PPT抗性筛选、PCR、PCRSouthern杂交检测获得IL-4阳性甘蓝植株，表明IL-4基因已转入甘蓝受体中。经ELISA和Western检测，说明IL-4基因已整合到甘蓝基因组，但表达量很低（0.23~281 ng/mL），且各植株间的表达量差异极大。
- Abstract:
- Taking the cabbage inbred lines seeds of transgenic IL-4 T0 materials named ‘92-1012-3-11’ as experimental material，the experiment researched the genetic transformation of IL-4gene into cabbage,which was mediated by Agrobacterium tumefaciens.The results showed that 396 strains of PPT resistant regenerated plants,137 strains of PCR positive plants,wherein the cotyledons with petiole conversion rate of 8.1%,hypocotyls differentiation rate was 5.8%.IL-4 positive cabbage plant in the positive transgenic line T0generation were detected by PPT resistance,PCR,PCRSouthern hybridization,and was showed that the IL-4 gene had been transferred into cabbage receptor.IL-4 gene had been integrated into the genome of cabbage which were detected by ELISA and Western detection,but it expressed very low (0.23~2.81 ng/mL),and there were difference greatly between the expression of each plant.
参考文献/References:
[1]MarquetBlouin E,Bouche E B,Steinmet A,et al.Neutralizing immunogenicity of transgenic carrot (Daucus carota L.)derived measles virus hemagglutinin[J].Plant Mol Biol,2003,51(4):459-469.
[2]Kumar G B,Ganapathi T R,Srinivas,et al.Secretion of hepatitis B surface antigen in transformed tobacco cell suspension cultures[J].Biotechnol Lett,2005,27(13):927-932.
[3]Tregoning J S,Nixon P,Kuroda H,et al.Expression of tetanus toxin fragment C in tobacco chloroplasts[J].Nucleic Acids Res,2003,31(4)：1174-1179.
[4]Askari M D,Tsao M S,Schuller H M,et al.The tobacco specific carcinogen,4(methylnitrosamino)1(3pyridyl)1butanone stimulates proliferation of immortalized human pancreatic duct epithelia through beta adrenergic transactivation of EGF receptors[J].J Cancer Res Clin Oncol,2005,131(10):639-648.
[5]Staub J M,Garcia B,Graves J,et al.High yield production of a human therapeutic protein in tobacco chloroplasts [J].Nat Biotechnol,2000,18(3)：333-338.
[6]龚剑,刘春林,郭纯,等.人胰岛素链基因表达载体的构建与大豆遗传转化[J].江苏农业科学,2005(2):27-31. [7]Bulat K L,Etokebe G E,Knezevic J,et al.Interferon gamma receptor 1 gene promoter polymorphisms(G-611A;T-56C) and susceptibility to tuberculosis[J].Scand J Immunol,2006,63(2):142-150.
[8]Conlon K,Osborne J,Morimoto C,et al.Comparison of lymphokine secretion and mRNA expression in the CD45RA+and CD45RO+subsets of human peripheral blood CD4+and CD8+ lymphocytes[J].Eur J Immunol,1995,25：644-648.
[9]Grimm E A,Mazumder A,Zhang H Z,et al.Lymphokine activated killer cell phenomenon：lysis of natural killer[J].J Exp Med,1982,155:1823-1841.
[10]Henney C S,Gall O K.Interleukin2 augments natural killer cell activity [J].Nature,1981,291:335-338. [11]王关林,方宏筠.植物基因工程[M].北京:科学出版社,2002:654.
[12]华志华,黄大年.转基因植物中外源基因的遗传学行为[J].植物学报,1999,41(1):1-5.
[13]贾庆利.外源基因在拟南芥转基因植株中的遗传研究[D].杨凌:西北农林科技大学,2004:24-26.
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备注/Memo
第一作者简介：金晓霞（1980-），女，博士，讲师，现主要从事植物分子生物学研究工作。E-mail：xiaoxia6195@126.com.
责任作者：于丽杰（1961-），女，博士，教授，硕士生导师，现主要从事植物基因工程研究工作。E-mail：yulijie1961@126.com.
基金项目：黑龙江省自然科学基金资助项目（C0014）。
收稿日期：2013-04-12