Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (2): 223-232.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0989
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REN Yan-jing1,2(), ZHANG Lu-gang3, ZHAO Meng-liang1, LI Jiang1,2, SHAO Deng-kui1,2()
Received:
2023-10-20
Online:
2024-02-26
Published:
2024-03-13
Contact:
SHAO Deng-kui
E-mail:renyan0202@163.com;2006990015@qhu.edu.cn
REN Yan-jing, ZHANG Lu-gang, ZHAO Meng-liang, LI Jiang, SHAO Deng-kui. cDNA Yeast Library Construction of Chinese Cabbage Seeds and Screening and Analysis of BrTTG1 Interacting Proteins[J]. Biotechnology Bulletin, 2024, 40(2): 223-232.
Fig. 1 Construction and identification of library A: Identification of the capacity of the primary library. B: Identification of inserts in the primary library. C: Identification of the capacity of the secondary library.D: Identification of inserts in the secondary library. E: Identification of the capacity of the yeast two-hybrid library. F: Identification of inserts in the yeast two-hybrid library
Fig. 2 Self-activation identification of pGBKT7-TTG1 bait vector A: Positive control. B: Negative control. C: SD/-Leu/-Trp/X-α-gal growth of gal medium. D: SD/-Leu/-Trp/X-α-gal growth of gal medium. E: SD/-Leu/-Trp/X-α-gal growth on gal/AbA medium
Fig. 3 Blue colonies screened by BrTTG1 in SD/-Leu/-Trp/-His/-Ade/X-α-gal/AbA(QDO/X/A)medium “-” represent negative control; “+” represent positive control
克隆编号Clone No. | 基因注释Gene annotation | 登录号GenBank access No. | 基因ID Gene ID |
---|---|---|---|
1 | Lipoamide dehydrogenase precursor, partial[Arabidopsis thaliana] | BAD95420.1 | BraA06g004870.3C |
2 | Hypothetical protein BRARA_C02124[Brassica rapa] | RID70072.1 | BraA03g021980.3C |
3 | Multiple organellar RNA editing factor 2, chloroplastic[B. rapa] | XP_009143860.1 | BraA05g011400.3C |
5 | Transcription factor MYB44[B. napus] | NP_001303071.1 | BraA01g001590.3C |
6 | DNA polymerase zeta processivity subunit[B. rapa] | XP_009117755.1 | BraA09g058190.3C |
7 | SNF1-related protein kinase regulatory subunit beta-2 isoform X1[B. napus] | XP_013655743.1 | BraA08g011810.3C |
10 | Protein phosphatase 2C 37[B. rapa] | XP_009146731.2 | BraA05g035520.3C |
12 | Hypothetical protein F2Q68_00033308, partial[B. cretica] | KAF2544242.1 | BraA03g030840.3C |
13 | ADP, ATP carrier protein 2, mitochondrial[B. rapa] | XP_009131345.2 | BraA03g005780.3C |
17 | Hypothetical protein F2Q69_00061138, partial[B. cretica] | KAF3574004.1 | BraA07g015680.3C |
24 | Hypothetical protein BRARA_I05150[B. rapa] | RID48647.1 | BraA09g061120.3C |
25 | Hypothetical protein F2Q70_00009385[B. cretica] | KAF2614836.1 | BraA09g001710.3C |
27 | Hypothetical protein BRARA_A02089[B. rapa] | RID79344.1 | BraA01g022400.3C |
31 | Unnamed protein product[B. rapa] | VDC79045.1 | BraA03g009070.3C |
33 | BnaA01g23700D[B. napus] | CDY18109.1 | BraA01g030470.3C |
39 | Unnamed protein product[B. oleracea] | VDD24561.1 | BraA02g028520.3C |
40 | Scarecrow-like protein 8[B. rapa] | XP_009127236.1 | BraA02g015300.3C |
43 | LOW QUALITY PROTEIN: tripeptidyl-peptidase 2-like[B. rapa] | XP_009137154.1 | BraA01g011620.3C |
44 | BnaC08g42880D[B. napus] | CDY22654.1 | BraA09g062530.3C |
45 | BnaC03g15020D[B. napus] | CDY38747.1 | BraA03g014440.3C |
46 | Transcription initiation factor IIE subunit beta isoform X2[B. rapa] | XP_009128687.1 | BraA02g028520.3C |
47 | Hypothetical protein BRARA_F02580[B. rapa] | RID59345.1 | BraA06g030810.3C |
48 | Hypothetical protein BRARA_G03388[B. rapa] | RID56172.1 | BraA07g039370.3C |
50-53 | COP9 signalosome complex subunit 5a[B. rapa] | XP_009115580.1 | BraA09g040180.3C |
54 | Thioredoxin M2, chloroplastic-like isoform X1[B. napus] | XP_013642642.1 | BraA09g002600.3C |
55 | E3 ubiquitin-protein ligase RGLG2[B. rapa] | XP_009121662.1 | BraA10g024620.3C |
56 | Uncharacterized protein LOC103841884[B. rapa] | XP_009116717.1 | BraA09g050150.3C |
57 | Hypothetical protein BRARA_E01984[B. rapa] | RID62948.1 | BraA05g024640.3C |
59 | Probable pectin methylesterase CGR2[B. rapa] | XP_009149708.1 | BraA06g017430.3C |
62 | E3 ubiquitin-protein ligase MBR2[B. rapa] | XP_009104796.1 | BraA07g028570.3C |
64 | Chitinase-like protein 1 isoform X1[B. rapa] | XP_009111023.1 | BraA08g034170.3C |
65 | GDSL esterase/lipase At1g71250, partial[B. rapa] | XP_009105797.3 | BraA07g035870.3C |
67 | PREDICTED: aconitate hydratase 1-like[Raphanus sativus] | XP_018463972.1 | BraA08g020550.3C |
68 | Hypothetical protein BRARA_J00468[B. rapa] | RID40424.1 | BraA10g005490.3C |
70 | NADPH-dependent oxidoreductase 2-alkenal reductase[B. rapa] | XP_009130330.1 | BraA05g041630.3C |
73 | SNF1-related protein kinase regulatory subunit beta-2 isoform X1[B. napus] | XP_013655743.1 | BraA08g011810.3C |
76 | Catalase-3 isoform X1[B. rapa] | XP_009149539.1 | BraA07g015680.3C |
82 | Mitogen-activated protein kinase 7[B. napus] | NP_001303162.1 | BraA07g002560.3C |
Table 1 pGBKT7-TTG1 interaction protein annotation
克隆编号Clone No. | 基因注释Gene annotation | 登录号GenBank access No. | 基因ID Gene ID |
---|---|---|---|
1 | Lipoamide dehydrogenase precursor, partial[Arabidopsis thaliana] | BAD95420.1 | BraA06g004870.3C |
2 | Hypothetical protein BRARA_C02124[Brassica rapa] | RID70072.1 | BraA03g021980.3C |
3 | Multiple organellar RNA editing factor 2, chloroplastic[B. rapa] | XP_009143860.1 | BraA05g011400.3C |
5 | Transcription factor MYB44[B. napus] | NP_001303071.1 | BraA01g001590.3C |
6 | DNA polymerase zeta processivity subunit[B. rapa] | XP_009117755.1 | BraA09g058190.3C |
7 | SNF1-related protein kinase regulatory subunit beta-2 isoform X1[B. napus] | XP_013655743.1 | BraA08g011810.3C |
10 | Protein phosphatase 2C 37[B. rapa] | XP_009146731.2 | BraA05g035520.3C |
12 | Hypothetical protein F2Q68_00033308, partial[B. cretica] | KAF2544242.1 | BraA03g030840.3C |
13 | ADP, ATP carrier protein 2, mitochondrial[B. rapa] | XP_009131345.2 | BraA03g005780.3C |
17 | Hypothetical protein F2Q69_00061138, partial[B. cretica] | KAF3574004.1 | BraA07g015680.3C |
24 | Hypothetical protein BRARA_I05150[B. rapa] | RID48647.1 | BraA09g061120.3C |
25 | Hypothetical protein F2Q70_00009385[B. cretica] | KAF2614836.1 | BraA09g001710.3C |
27 | Hypothetical protein BRARA_A02089[B. rapa] | RID79344.1 | BraA01g022400.3C |
31 | Unnamed protein product[B. rapa] | VDC79045.1 | BraA03g009070.3C |
33 | BnaA01g23700D[B. napus] | CDY18109.1 | BraA01g030470.3C |
39 | Unnamed protein product[B. oleracea] | VDD24561.1 | BraA02g028520.3C |
40 | Scarecrow-like protein 8[B. rapa] | XP_009127236.1 | BraA02g015300.3C |
43 | LOW QUALITY PROTEIN: tripeptidyl-peptidase 2-like[B. rapa] | XP_009137154.1 | BraA01g011620.3C |
44 | BnaC08g42880D[B. napus] | CDY22654.1 | BraA09g062530.3C |
45 | BnaC03g15020D[B. napus] | CDY38747.1 | BraA03g014440.3C |
46 | Transcription initiation factor IIE subunit beta isoform X2[B. rapa] | XP_009128687.1 | BraA02g028520.3C |
47 | Hypothetical protein BRARA_F02580[B. rapa] | RID59345.1 | BraA06g030810.3C |
48 | Hypothetical protein BRARA_G03388[B. rapa] | RID56172.1 | BraA07g039370.3C |
50-53 | COP9 signalosome complex subunit 5a[B. rapa] | XP_009115580.1 | BraA09g040180.3C |
54 | Thioredoxin M2, chloroplastic-like isoform X1[B. napus] | XP_013642642.1 | BraA09g002600.3C |
55 | E3 ubiquitin-protein ligase RGLG2[B. rapa] | XP_009121662.1 | BraA10g024620.3C |
56 | Uncharacterized protein LOC103841884[B. rapa] | XP_009116717.1 | BraA09g050150.3C |
57 | Hypothetical protein BRARA_E01984[B. rapa] | RID62948.1 | BraA05g024640.3C |
59 | Probable pectin methylesterase CGR2[B. rapa] | XP_009149708.1 | BraA06g017430.3C |
62 | E3 ubiquitin-protein ligase MBR2[B. rapa] | XP_009104796.1 | BraA07g028570.3C |
64 | Chitinase-like protein 1 isoform X1[B. rapa] | XP_009111023.1 | BraA08g034170.3C |
65 | GDSL esterase/lipase At1g71250, partial[B. rapa] | XP_009105797.3 | BraA07g035870.3C |
67 | PREDICTED: aconitate hydratase 1-like[Raphanus sativus] | XP_018463972.1 | BraA08g020550.3C |
68 | Hypothetical protein BRARA_J00468[B. rapa] | RID40424.1 | BraA10g005490.3C |
70 | NADPH-dependent oxidoreductase 2-alkenal reductase[B. rapa] | XP_009130330.1 | BraA05g041630.3C |
73 | SNF1-related protein kinase regulatory subunit beta-2 isoform X1[B. napus] | XP_013655743.1 | BraA08g011810.3C |
76 | Catalase-3 isoform X1[B. rapa] | XP_009149539.1 | BraA07g015680.3C |
82 | Mitogen-activated protein kinase 7[B. napus] | NP_001303162.1 | BraA07g002560.3C |
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