甜菜BvM14-STPK蛋白激酶互作蛋白的鉴定及筛选

doi:10.11924/j.issn.1000-6850.casb20190600323

中国农学通报 ›› 2020, Vol. 36 ›› Issue (1): 30-34.doi: 10.11924/j.issn.1000-6850.casb20190600323

所属专题：生物技术；园艺

甜菜BvM14-STPK蛋白激酶互作蛋白的鉴定及筛选

吕家宝¹^,², 余梦頔¹^,², 李海英¹^,²()

¹ 黑龙江大学农业微生物技术教育部工程研究中心,哈尔滨 150500;
² 黑龙江大学生命科学学院/黑龙江省普通高校分子生物学重点实验室,哈尔滨 150080

收稿日期:2019-06-24 修回日期:2019-07-25 出版日期:2020-01-05 发布日期:2020-01-07
通讯作者: 李海英
作者简介:吕家宝,男,1995年出生,河南禹州人,硕士研究生,研究方向：植物分子生物学。通信地址：150080 黑龙江省哈尔滨市南岗区学府路74号黑龙江大学生命科学学院320室,E-mail：867969369@qq.com。
基金资助:
国家自然科学基金“甜菜M14品系抗氧化酶系统响应盐胁迫应答过程的研究”(31471552);黑龙江省高校创新团队建设计划项目“寒区植物重要基因资源的挖掘与种质创新”(2014TD004);黑龙江大学2019年研究生创新科研资金项目“BvM14-STPK蛋白激酶互作蛋白的筛选与互作关系验证”(YJSCX2019-202HLJU)

BvM14-STPK Protein Kinase Interaction Protein in Beta vulgaris: Identification and Screening

Lv Jiabao¹^,², Yu Mengdi¹^,², Li Haiying¹^,²()

¹ Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education, Heilongjiang University, Harbin 150500;
² Key Laboratory of Molecular Biology, College of Heilongjiang Province, School of Life Sciences, Heilongjiang University, Harbin 150080

Received:2019-06-24 Revised:2019-07-25 Online:2020-01-05 Published:2020-01-07
Contact: Li Haiying

摘要/Abstract

摘要：

为进一步探究BvM14-STPK蛋白激酶在甜菜中应答盐胁迫的信号途径,本研究对BvM14-STPK蛋白激酶进行了互作蛋白的鉴定及筛选。使用转基因技术获得转基因烟草,使用亲和纯化串联质谱技术鉴定蛋白复合物,通过搜索烟草蛋白质数据库,获得BvM14-STPK蛋白激酶的候选互作蛋白;使用实验室甜菜M14品系盐胁迫转录组数据,对得到的互作蛋白的编码基因进行盐胁迫下的转录水平分析。结果表明,在烟草数据库中,非盐处理条件下获得3个BvM14-STPK蛋白候选互作蛋白质,盐处理条件下获得6个候选互作蛋白质;经过盐胁迫转录组数据分析,发现有4个互作蛋白编码基因在不同的组织部位、不同的盐浓度条件下应答盐胁迫。为后续进一步在植物体内验证BvM14-STPK蛋白激酶与候选互作蛋白的相互作用及甜菜抗盐机理奠定良好的基础。

关键词: 甜菜M14品系, 蛋白激酶, 亲和纯化串联质谱, 互作蛋白, 盐胁迫

Abstract:

To further explore the signal pathway of BvM14-STPK protein kinase in response to salt stress in sugar beet, the interaction proteins of BvM14-STPK protein kinase were identified and screened in this study. Transgenic tobacco was obtained by transgenic technology, and the protein complex was identified by affinity purification tandem mass spectrometry (TAP/MS). The candidate interaction proteins of BvM14-STPK protein kinase were obtained by searching the tobacco protein database, and the RNA-sequencing data of sugar beet M14 was used to analyze the transcription level of the genes which encoded the candidate interaction proteins under salt stress. The results showed that, three BvM14-STPK protein candidate interaction proteins were obtained under non-salt treatment, and six candidate interaction proteins were obtained under salt treatment in the tobacco database. The transcriptome data analysis under salt stress showed that four interaction protein-coding genes responded to salt stress at different tissues and different salt concentrations. It is a positive foundation for further verification of the interaction between BvM14-STPK protein kinase and its candidate interaction proteins in plants and the salt resistance mechanism of sugar beet.

Key words: sugar beet M14, protein kinase, TAP/MS, protein-protein interaction, salt stress

中图分类号:

S566.3

吕家宝, 余梦頔, 李海英. 甜菜BvM14-STPK蛋白激酶互作蛋白的鉴定及筛选[J]. 中国农学通报, 2020, 36(1): 30-34.

Lv Jiabao, Yu Mengdi, Li Haiying. BvM14-STPK Protein Kinase Interaction Protein in Beta vulgaris: Identification and Screening[J]. Chinese Agricultural Science Bulletin, 2020, 36(1): 30-34.

图/表 4

参考文献 21

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Accession		Description	Coverage	#Peptides	# Unique Peptides	#AAs	MW/kDa	pI
XP_009771888.1	AB	PREDICTED: heat shock 70 kDa protein-like [Nicotiana sylvestris]	3.490136571	2	2	659	72.292	5.39
XP_016439103.1	AB	PREDICTED: DNA repair protein RAD16-like [Nicotiana tabacum]	1.187904968	2	2	926	104.76	8.22
XP_016437499.1	AB	PREDICTED: chaperone protein ClpB1 [Nicotiana tabacum]	3.951701427	4	4	911	101.35	6.21
XP_016475992.1	B	PREDICTED: probable serine/ threonine-protein kinase At1g54610 [Nicotiana tabacum]	2.456140351	2	2	570	65.306	9.61
XP_009775590.1	B	PREDICTED: zinc finger BED domain-containing protein RICESLEEPER 1-like, partial [Nicotiana sylvestris]	1.431980907	2	2	838	94.175	7.93
XP_016495333.1	B	PREDICTED: L-type lectin-domain containing receptor kinase S.4-like [Nicotiana tabacum]	2.026049204	2	2	691	77.049	6.68

Accession		Description	Coverage	#Peptides	# Unique Peptides	#AAs	MW/kDa	pI
XP_009771888.1	AB	PREDICTED: heat shock 70 kDa protein-like [Nicotiana sylvestris]	3.490136571	2	2	659	72.292	5.39
XP_016439103.1	AB	PREDICTED: DNA repair protein RAD16-like [Nicotiana tabacum]	1.187904968	2	2	926	104.76	8.22
XP_016437499.1	AB	PREDICTED: chaperone protein ClpB1 [Nicotiana tabacum]	3.951701427	4	4	911	101.35	6.21
XP_016475992.1	B	PREDICTED: probable serine/ threonine-protein kinase At1g54610 [Nicotiana tabacum]	2.456140351	2	2	570	65.306	9.61
XP_009775590.1	B	PREDICTED: zinc finger BED domain-containing protein RICESLEEPER 1-like, partial [Nicotiana sylvestris]	1.431980907	2	2	838	94.175	7.93
XP_016495333.1	B	PREDICTED: L-type lectin-domain containing receptor kinase S.4-like [Nicotiana tabacum]	2.026049204	2	2	691	77.049	6.68

甜菜BvM14-STPK蛋白激酶互作蛋白的鉴定及筛选

BvM14-STPK Protein Kinase Interaction Protein in Beta vulgaris: Identification and Screening

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