[1]欧阳秋玲,刘士德,张建华,等.采用酵母双杂交法筛选PSRPK相关蛋白基因[J].深圳大学学报理工版,2006,23(3):222-229.
 OUYANG Qiu-ling,LIU Shi-de,ZHANG Jian-hua,et al.Screening of the psrpk一related protein genes with yeast two-hybrid method[J].Journal of Shenzhen University Science and Engineering,2006,23(3):222-229.
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采用酵母双杂交法筛选PSRPK相关蛋白基因()
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《深圳大学学报理工版》[ISSN:1000-2618/CN:44-1401/N]

卷:
第23卷
期数:
2006年3期
页码:
222-229
栏目:
生命科学
出版日期:
2006-07-30

文章信息/Info

Title:
Screening of the psrpk一related protein genes with yeast two-hybrid method
文章编号:
1000-2618(2000)03-0222-08
作者:
欧阳秋玲刘士德张建华王绍文田生礼邢苗
深圳大学生命科学学院;深圳市微生物基因工程重点实验室,深圳518060
Author(s):
OUYANG Qiu-ling LIU Shi-deZHANG Jian-huaWANG Shao-wen TIAN Sheng-liand XING Miao
College of Life Science Shenzhen Key Laboratory of Microbial Gene Engineering Shenzhen University
关键词:
多头绒胞菌酵母双杂交PSRPKPlasmin C BCAT类似蛋白MSF1类似蛋白mixedlinked glucanase precursor类似蛋白FCY1p类似蛋白40S ribosomal protein S2 类似蛋白
Keywords:
Pltysarum polycephalum Two-hybrid system PSRPK Plasmin C BCAT like protein MSF1-like protein mixed-linked glucanase precursor like protein FCY1p like protein 4OS ribosomal protein S2 like protein
分类号:
Q2
文献标志码:
A
摘要:
本课题组在前期研究中,从多头绒胞菌中分离到一个SR蛋白激酶基因psrpk,并将其编码蛋白命名为PSRPK(SR protein kinase of Physarum Polycephalum).为分离PSRPK相关蛋白基因以了解PSRPK的功能,构建了转化率为2×106转化子/3μg pGADT7-Rec、密度为5.35×108cells/mL、滴度为2.34× 109cfu/mL的多头绒泡菌酵母双杂交AD库.以PSRPK为饵蛋白筛选该文库得到接合率为41.18%的杂交酵母在SD/-Leu/-Trp/-Ade/-His培养板上筛选获得1476个杂交克隆,其中,X-gal滤纸显色呈强蓝色的克隆有342个。对大于500bp的67个克隆测序获得35个cDNA 片段,其中编码plasmin C, branched-chain amino acid aminotransferase (BCAT)类似蛋白、MSFI类似蛋白、mixed-linked glucanase precursor类似蛋白、FCY1p类似蛋白和40S ribosomal protein S2类似蛋白等7个cDNA片段在阳性杂交酵母克隆中出现多次,其余仅出现一次。在35个cDNA片段的编码序列中有15个具有同源蛋白,31个编码序列的丝氨酸含量大于或等于6%,符合激酶底物的组成特征。
Abstract:
A novel SR protein-specific kinases(SRPKs) gene psrpk,,was recently isolated from (SR protein kinase of Physarum polycephalum).To investigate the biological function of PSRPK and screen for the PSRPK-related proteins, a and the coding protein was the biological function of PSRPK and screen for the PSRPK-related proteins, a 2 x106 transformants/3μg pGADT7-Rec of transformation efficiency,5. 35 x 10R cells/ml, of cell density and 2. 34× 109 cfu/mL titre of P. Pnlycephalum two-hybrid AD library was constructed.When screening the AD library with psrpk, 1476 diploid yeast clones on the SD/-Leu/-Trp/-Adc/一His plates were obtained with a mating efficiency of 41.8%. After screening on X-gal filter, 342 positive clones were confirmed with blue color,67 of them were sequenced basing on the inserts size(≥500bp),35 cDNA fragments were obtained. Among the cDNA fragments, 7 of them were presented in multiclones, which encode the plasmin C, branched-chain amino acid aminotransferase (BCAT)like protein,MSF1-like protein,mixed-linked glucanase precursor like protein, FCY1p like protein and 4OS ribosomal protein S2 like proteins of other species. Most of the coding proteins contain more than 6% serine, showing a character of the substrates of the kinases.

相似文献/References:

[1]冯波,曾虹燕,钟英丽,等.转录因子AP-2α与TES蛋白相互作用分析[J].深圳大学学报理工版,2008,25(4):432.
 FENG Bo,ZENG Hong-yan,ZHONG Ying-li,et al.Analysis of interaction of AP-2α and TES[J].Journal of Shenzhen University Science and Engineering,2008,25(3):432.

更新日期/Last Update: 2015-06-26