Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (9): 1723-1732.DOI: 10.3969/j.issn.1000-2561.2020.09.001
• Omics & Biotechnology • Next Articles
ZHANG Yuhang1,2,*,PAN Ranran1,*,LI Fei1,TAO Zhiqiang1,2,WANG Ying1,GAO Heqiong1,ZHUANG Nansheng1,**()
Received:
2019-09-09
Revised:
2019-12-23
Online:
2020-09-25
Published:
2020-10-16
Contact:
ZHANG Yuhang,PAN Ranran,ZHUANG Nansheng
CLC Number:
ZHANG Yuhang,PAN Ranran,LI Fei,TAO Zhiqiang,WANG Ying,GAO Heqiong,ZHUANG Nansheng. Cloning, Physical Location and Expression Analysis of HbGRF in Hevea brasiliensis[J]. Chinese Journal of Tropical Crops, 2020, 41(9): 1723-1732.
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基因 Gene | 上游引物(5ʹ-3ʹ) Forward primer (5ʹ-3ʹ) | 下游引物(5ʹ-3ʹ) Reverse primer (5ʹ-3ʹ) | 用途 Use |
---|---|---|---|
HbGRF1 | AAATAGTTACAAAGCAGGC | TTCTTCAGTGTAGAGGTGGTG | 基因克隆 |
GCTGCCTCTTCCTCATC | AACACCTTGGACTACGA | 基因定位 | |
AGAAAGCCTGTGGAAGGTCA | CCACCACAGCTGCTGAAATA | 定量分析 | |
HbGRF2 | AAGTTGCTGGTAAGAACAAG | AAAGATGCCAAAACTCCTCCT | 基因克隆 |
GCACCCTTTATGTTTCC | CAGCAAGATTTGTTAGTAGAG | 基因定位 | |
CAGGTACATGTTGGCTGGTG | ATACGGTGGAGAGCGAAGAA | 定量分析 | |
HbGRF3 | TCACTACCGTCCGTTTCCATC | CAACCACGGCAATCCAATC | 基因克隆 |
ACCGCATAATAACCAAT | GTTCAAGAAAGCCTGTG | 基因定位 | |
AGATCCAAATGGGGTTAGGC | GTGTAGCCCCTGTCCAAGAA | 定量分析 | |
18S rRNA | GCTCGAAGACGATCAGATACC | TTCAGCCTTGCGACCATAC | 定量分析 |
Tab. 1 Primers sequences for PCR
基因 Gene | 上游引物(5ʹ-3ʹ) Forward primer (5ʹ-3ʹ) | 下游引物(5ʹ-3ʹ) Reverse primer (5ʹ-3ʹ) | 用途 Use |
---|---|---|---|
HbGRF1 | AAATAGTTACAAAGCAGGC | TTCTTCAGTGTAGAGGTGGTG | 基因克隆 |
GCTGCCTCTTCCTCATC | AACACCTTGGACTACGA | 基因定位 | |
AGAAAGCCTGTGGAAGGTCA | CCACCACAGCTGCTGAAATA | 定量分析 | |
HbGRF2 | AAGTTGCTGGTAAGAACAAG | AAAGATGCCAAAACTCCTCCT | 基因克隆 |
GCACCCTTTATGTTTCC | CAGCAAGATTTGTTAGTAGAG | 基因定位 | |
CAGGTACATGTTGGCTGGTG | ATACGGTGGAGAGCGAAGAA | 定量分析 | |
HbGRF3 | TCACTACCGTCCGTTTCCATC | CAACCACGGCAATCCAATC | 基因克隆 |
ACCGCATAATAACCAAT | GTTCAAGAAAGCCTGTG | 基因定位 | |
AGATCCAAATGGGGTTAGGC | GTGTAGCCCCTGTCCAAGAA | 定量分析 | |
18S rRNA | GCTCGAAGACGATCAGATACC | TTCAGCCTTGCGACCATAC | 定量分析 |
名称 Name | 分子式 Molecular formula | 氨基酸Amino acid | 分子量 Molecular weight/kDa | 等电点 pI | 蛋白质不稳定指数 Protein instability index | 脂肪 系数 Fat coefficient | 疏水性 Hydrophobicity | 亚细胞定位 Subcellular localization | 起始密 码子 Start codon | 终止密 码子 Stop codon | |
---|---|---|---|---|---|---|---|---|---|---|---|
数值 Value | 评价 Evaluate | ||||||||||
GRF1 | C2279H3540 N668O745S20 | 609 | 65.663 | 8.48 | 51.08 | 不稳定 | 69.03 | -0.541 | 细胞核 | ATG | TAA |
GRF2 | C1798H2806 N524O565S12 | 384 | 41.188 | 8.77 | 56.06 | 不稳定 | 62.03 | -0.580 | 细胞核 | ATG | TGA |
GRF3 | C2279H3540 N668O745S20 | 494 | 52.858 | 6.09 | 51.27 | 不稳定 | 57.98 | -0.582 | 细胞核 | ATG | TAG |
Tab. 2 Analysis of composition and physicochemical properties of HbGRF gene proteins sequence
名称 Name | 分子式 Molecular formula | 氨基酸Amino acid | 分子量 Molecular weight/kDa | 等电点 pI | 蛋白质不稳定指数 Protein instability index | 脂肪 系数 Fat coefficient | 疏水性 Hydrophobicity | 亚细胞定位 Subcellular localization | 起始密 码子 Start codon | 终止密 码子 Stop codon | |
---|---|---|---|---|---|---|---|---|---|---|---|
数值 Value | 评价 Evaluate | ||||||||||
GRF1 | C2279H3540 N668O745S20 | 609 | 65.663 | 8.48 | 51.08 | 不稳定 | 69.03 | -0.541 | 细胞核 | ATG | TAA |
GRF2 | C1798H2806 N524O565S12 | 384 | 41.188 | 8.77 | 56.06 | 不稳定 | 62.03 | -0.580 | 细胞核 | ATG | TGA |
GRF3 | C2279H3540 N668O745S20 | 494 | 52.858 | 6.09 | 51.27 | 不稳定 | 57.98 | -0.582 | 细胞核 | ATG | TAG |
Fig. 1 Prediction of secondary structure of HbGRF proteins in H. brasiliensis Red stripe: spiral structure; Blue stripe: chain; Yellow stripe: hidden structure; Green stripe: irregular structure; Red small square: protein binding area; Yellow small circle: DNA binding area; Purple small circle: RNA binding area; Gray small circle: nucleotide binding area.
Fig. 4 IS-PCR with HbGRF1 at chromosomes on different phase of mitosis of ‘Reyan 7-33-97’ A: Interphase cell; B: Prometaphase cell; C: Metaphase cell; D: Negative control; E-F: Metaphase cell division and its karyotype; Arrows indicated the signal of HbGRF1.
Fig. 5 IS-PCR with HbGRF2 at chromosomes on different phase of mitosis of ‘Reyan 7-33-97’ A: Interphase cell; B: Prometaphase cell; C: Metaphase cell; D: Negative control; E-F: Metaphase cell division and its karyotype; Arrows indicated the signal of HbGRF2.
Fig. 6 FISH with HbGRF3 at chromosomes on different phase of mitosis of ‘Reyan 7-33-97’ A: Interphase cell; B: Prometaphase cell; C: Metaphase cell; D: Negative control; E-F: Metaphase cell division and its karyotype; Arrows indicated the signal of HbGRF3.
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