当前位置: 首页 > 文章 > 将IL-2基因构建到叶绿体转化载体并转入烟草 江苏农业学报 2006,22 (4) 379-383
Position: Home > Articles > Transformation of Human IL-2 Gene into Tobacco by Construction of A Chloroplast Vector Jiangsu Journal of Agricultural Sciences 2006,22 (4) 379-383

将IL-2基因构建到叶绿体转化载体并转入烟草

作  者:
黄益洪;刘蔼民;汤日圣;张大栋;童红玉;朱鑫泉;沙钟瑞;吴光南
单  位:
江苏省农业科学院食品质量安全与检测研究所;中国农业生物技术学会;江苏省农业科学院农业生物遗传生理研究所
关键词:
烟草;叶绿体转化;壮观霉素;人白细胞介素2基因
摘  要:
以人白细胞介素2(IL-2)基因为目的基因,烟草accD和ORF184同源序列为侧翼引导序列构建成烟草叶绿体转化载体pAIL。以大烟草NC89为转化受体,通过基因枪方法对烟草叶片进行轰击转化。轰击参数为:每枪0.6μm的金粉300~500μg,轰击距离6 cm,真空度9.48×104Pa。在500 mg/L的壮观霉素筛选压下,经约30 d的筛选分化,逐渐产生绿色的抗壮观霉素芽或愈伤,目前已获得31株(丛)抗性芽及8块抗性愈伤。用IL-2基因的PCR引物对4株较大的小苗进行检测,初步证明3株为转基因植株。
译  名:
Transformation of Human IL-2 Gene into Tobacco by Construction of A Chloroplast Vector
作  者:
HUANG Yi-hong~1,LIU Ai-min~2,TANG Ri-sheng~1,ZHANG Da-dong~1,TONG Hong-yu~1,ZHU Xin-quan~3,SHA Zhong-rui~3,WU Guang-nan~1 (1.Institute of Agrobiological Genetic and Physiology,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;2.Institute of Food Safety Research,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China;3.Chinese Society of Agricultural Biotechnology,Beijing 100026,China)
关键词:
tobacco;chloroplast transformation;spectinomycin;IL-2 gene
摘  要:
A vector containing human interleukin-2(IL-2) gene was constructed with accD and ORF184 sequences of tobacco chloroplast as flanking sequences.This vector was used to transform IL-2 gene into tobacco NC89(Nictiana tabacum) by a particle bombardment system.The main parameters include Au particle(0.6 μm) 300-500 μg per shot,bombardment distance 6 cm,and 9.48×10~4 Pa.Hg vaccum.The spectinomycin resistant plants were selected on MS.S medium supplemented with 500 mg/L spectinomycin.Some green spec-resistant plantlets or calli appeared after about a month through bombardment.So far,31 spec-resistant plantlets and 8 calli were obtained.Three out of four bigger plantlets were proved as transgenic plants by PCR assay with IL-2 gene.

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