期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

同源重组快速构建甘蓝SCR酵母双杂交诱饵载体及其自激活作用的检测

Rapid Construction of Bait Vector Containing SCR from Brassica oleracea L.by Homologous Recombination and Evaluation of Its Self-Activation in the Yeast Two-hybrid System
原文传递
导出
摘要 采用巢式PCR获得甘蓝SCR的成熟肽编码区,利用同源重组技术首次将其克隆到pGBKT7载体中,构建酵母双杂交系统的诱饵载体pGBKT7-SCR,结果表明:通过巢式PCR获得了正确的甘蓝SCR成熟肽编码区,并成功构建到pGBKT7诱饵载体中,且转化有诱饵载体的Y2HGold在SD/-Trp营养缺陷平板上生长良好,而在SD/-His-Trp和SD/-Trp/X-a-Gal/AbA营养缺陷平板上皆不能生长,说明对报告基因无自激活作用;且毒性实验也表明,SCR蛋白对酵母没有毒害作用. The mature coding sequence of S-locus cysteine-rich gene（SCR） in Brassica oleracea L.was amplified by nested-PCR and cloned into the yeast plasmid pGBKT7 by homologous recombination.The recombinant bait plasmid pGBKT7-SCR was verified by sequencing before transformation into competent yeast cells.Sequence analysis showed that the SCR sequence was right and the yeast vector of pGBKT7-SCR was constructed correctly.The transformed Y2HGold Yeast（pGBKT7-SCR） could grow on the SD/-trp plates,while no colony could survive on the SD/-his-trp and SD/-Trp/X-a-Gal/AbA pate,indicating that the bait vector of pGBKT7-SCR would not self-activate the reporter gene.Furthermore,the recombinant bait plasmid showed no toxic effect on yeast Y2HGold cell.These results suggested that the recombinant bait plasmid pGBKT7-SCR can be used to study the interactions between SCR and SRK by the yeast two-hybrid system and lay a foundation for obtaining insight into the mechanism of self-incompatibility in B.oleracea L..
作者 罗兵 朱利泉 薛丽琰 张贺翠 彭一波 杨红 杨昆 高启国 李成琼 王小佳
机构地区 西南大学植物生理生物化学实验室 常熟理工学院生物与食品工程学院 西南大学重庆市蔬菜学重点实验室
出处 《西南大学学报（自然科学版）》 CAS CSCD 北大核心 2012年第8期79-85,共7页 Journal of Southwest University(Natural Science Edition)
基金 国家自然科学基金资助项目(30971849)
关键词 甘蓝 SCR蛋白 同源重组 酵母双杂交 诱饵载体 自激活 Brassica oleracea L. SCR protein homologous recombination yeast two-hybrid system bait plasmid self-activation
分类号 Q78 [生物学—分子生物学]
  • 相关文献

参考文献17

  • 1TAKAYAMA S, SHIMOSATO H, SHIBA H, et al. Direct Ligand-Receptor Complex Interaction Controls Brasszca Self-Incompatibility [J]. Nature, 2001, 413 : 534- 538.
  • 2SCHOPFER C R, NASRALLAH M E, NASRALLAH J B. The Male Determinant of Self-Incompatibility in Brassica[J]. Science, 1999, 286: 1697-1700.
  • 3SUZUKI G, KAI N, HIROSE T, et al. Genomic Organization of the S Locus: Identification and Characterization of Genes in SLG/SRK Region of $9 Haplotype of Brassica Campestris (syn. rapa) [J]. Genetics, 1999, 153:391- 400.
  • 4SHIBA H, TAKAYAMA S, IWANO M, et al. A Pollen Coat Protein, SP11/SCR, Determines the Pollen S-Specificity in the Self-Incompatibility of Brassica Species [J]. Plant Physiology, 2001, 125:2095-2103.
  • 5杨洋,高启国,宋明,牛义,汤青林,朱利泉,王小佳.甘蓝自交不亲和决定因子的体外表达和相互作用的检测[J].园艺学报,2009,36(3):355-362. 被引量：8
  • 6STEIN J C, HOWLETT B, BOYES D C, et al. Molecular Cloning of a Putative Receptor Protein Kinase Gene Encoded at the Self-Incomaptibility Locus of Brassica Oleracea [J]. Proc Natl Acad Sci USA, 1991, 88: 8816-8820.
  • 7TAKASAKI T, HATAKEYAMA K, SUZUKI G, et al. The S Receptor Kinase Determines Self-Incompatibility in Brassica Stigma [J]. Nature, 2000, 403: 913-916.
  • 8罗兵,薛丽琰,朱利泉,张贺翠,彭一波,陈松,杨红,杨昆,李成琼,王小佳.利用酵母双杂交法检测甘蓝SCR与SRK之间的相互作用[J].作物学报,2011,37(4):579-586. 被引量：6
  • 9KACHROO A, SCHOPFER C R, NASRALLAH M E, et al. Allele-Specific Receptor-Ligand Interactions in Brassica Self-Incompatibility [J]. Science, 2001, 293 : 1824- 1826.
  • 10HIROKO SHIMOSATO, NAOHIKO YOKOTA, HIROSHI SHIBA, et al. Characterization of the SP11/SCR High- Affinity Binding Site Involved in Self/Nonself Recognition in Brassica Self-Incompatibility [J]. Plant Cell, 2007, 19(1) : 107-117.

二级参考文献39

  • 1Cui Y, Bi Y M, Brugiere N, Amoldo M, Rothstein S J. 2000. The S locus glycoprotein and the S receptor kinase are sufficient for self-pollen rejection in Brassica. Proc Natl Acad Sci USA, 97:3713 -3717.
  • 2Franklin-Tong V E, Franklin C H. 2000. Self-incompatibility in Brassica: The elusive pollen S gene is identified. Plant Cell, 12:305 -308.
  • 3Goring D R. 2000. The search for components of the self-incompatibility signaling pathway (S) in Brassica napus. Annals of Botany, 85 : 171 - 179.
  • 4Kachroo A, Sehopfer C R, Nasrallah M E, Nasrallah J B. 2001. Allele-specific receptor-ligand interaction in Brassica self-incompatibilily. Science, 293 (5536) : 1824 - 1826.
  • 5Kao T H, McCubbin A G. 2000. Plant biology : A social stigma. Nature, 403 (6772) : 840 -841.
  • 6Schopfer C R, Nasrallah M E, Nasrallah J B. 1999. The male determinant of self-incompatibillty in Brassica. Science, 286 (5445): 1697 - 1700.
  • 7Shiba H, Takayama S, Iwano M, Shimosato H, Fonato M, Nakagawa T, Che F S, Suzuki G, Watanabe M, Hinata K, Isogai A. 2001. A pollen coat protein, SP11/SCR, determines the pollen S-specificity in the self-incompatibility of Brassica species. Plant Physiology 125:2095 - 2103.
  • 8Silva N F, Stone S L, Christie L N, Sulaman W, Nazarian K A P, Burentt L A, Arnoldo M A, Rothstein S J, Coring D R. 2001. Expression of the S receptor kinase ill self-compatible Brassica napus cv. Westar leads to the allele-specific rejection of self-incompatible Brassica napus pollen. Mol Genet Genom, 265:552 -559.
  • 9Stein J C, Howlett B, Boyes D C, Nasarllah M E, Nasarllah J B. 1991, Molecular cloning of a putative receptor protein kinase encoded at the self- incompatibility locus of Brassica oleracea. Proc Nat Acad Sci USA, 8:8816 -8820.
  • 10Suzuki G, Kai N, Hirose T. 1999. Genomic organization of the S locus: Identification and characterization of genes in SLG/SRK region of S^9 haplotype of Brassica campestris (syn. rapa). Genetics, 153 (1) : 391 -400.

共引文献12

西南大学学报（自然科学版）
2012年 第8期

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部