Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (4): 163-173.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1272
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MA Xin-xin(), XU Yang, ZHAO Huan-huan, HUO Zhao-yan, WANG Shu-bin, ZHONG Feng-lin()
Received:
2021-10-09
Online:
2022-04-26
Published:
2022-05-06
Contact:
ZHONG Feng-lin
E-mail:1126691073@qq.com;zhong591@fafu.edu.cn
MA Xin-xin, XU Yang, ZHAO Huan-huan, HUO Zhao-yan, WANG Shu-bin, ZHONG Feng-lin. Identification of Tomato 4CL Gene Family and Expression Analysis Under Nitrogen Treatment[J]. Biotechnology Bulletin, 2022, 38(4): 163-173.
类型 Type | 基因ID Gene ID | 基因名称Gene name | 正向引物Forward(5'-3') | 反向引物Reverse(5'-3') |
---|---|---|---|---|
RT-qPCR | Solyc03g097030 | Sl4CL01 | TTTGAGGAGAGGACATGTATTATCG | GACGATGCTAGAATGGTAAA |
Solyc03g117870 | Sl4CL02 | ATGTTGACACACAAAGGCT | ATTGCTGCTCCGACTCTC | |
Solyc06g068650 | Sl4CL03 | TGTCTTCCTTGGTGCGTC | GCATAATCCTTCACCTTGTTC | |
Solyc08g076300 | Sl4CL04 | TAAGTCAGCAAGATACAGCG | GAGAGAAAGCCCATACACAT | |
Solyc11g069050 | Sl4CL05 | TCAACCTATGAGTATCTTTGCTGC | AAAACCGTTGTGCCGTGC | |
Solyc12g042460 | Sl4CL06 | AGTAAGGATAGTCATTTCTGGG | GCTAAGCACATTGTTATAGGTG | |
Actin | GTCCTCTTCCAGCCATCCAT | ACCACTGAGCACAATGTTACCG | ||
RT-PCR | Solyc06g068650 | Sl4CL03 | ATGACAAATGCAACGATGGAG | ATTTGGA TATCCAGCAG CT |
pCAMBIA1302 | GTGGATTGATGTGATATCTCC | CTGACAGAAAATTTGTGCCC |
Table 1 Primers for PCR
类型 Type | 基因ID Gene ID | 基因名称Gene name | 正向引物Forward(5'-3') | 反向引物Reverse(5'-3') |
---|---|---|---|---|
RT-qPCR | Solyc03g097030 | Sl4CL01 | TTTGAGGAGAGGACATGTATTATCG | GACGATGCTAGAATGGTAAA |
Solyc03g117870 | Sl4CL02 | ATGTTGACACACAAAGGCT | ATTGCTGCTCCGACTCTC | |
Solyc06g068650 | Sl4CL03 | TGTCTTCCTTGGTGCGTC | GCATAATCCTTCACCTTGTTC | |
Solyc08g076300 | Sl4CL04 | TAAGTCAGCAAGATACAGCG | GAGAGAAAGCCCATACACAT | |
Solyc11g069050 | Sl4CL05 | TCAACCTATGAGTATCTTTGCTGC | AAAACCGTTGTGCCGTGC | |
Solyc12g042460 | Sl4CL06 | AGTAAGGATAGTCATTTCTGGG | GCTAAGCACATTGTTATAGGTG | |
Actin | GTCCTCTTCCAGCCATCCAT | ACCACTGAGCACAATGTTACCG | ||
RT-PCR | Solyc06g068650 | Sl4CL03 | ATGACAAATGCAACGATGGAG | ATTTGGA TATCCAGCAG CT |
pCAMBIA1302 | GTGGATTGATGTGATATCTCC | CTGACAGAAAATTTGTGCCC |
基因名称Gene name | 氨基酸个数 Number of amino acids | 分子量 Molecular weight/kD | 等电点 Isoelectric point/pI | 不稳定指数Instability index | 总平均亲水系数Grand average of hydropathicity | 原子总数Total number of atoms | 亚细胞定位Subcellular localization |
---|---|---|---|---|---|---|---|
Sl4CL01 | 570 | 62080.54 | 5.34 | 34.92 | 0.106 | 8820 | Peroxisome Chloroplast |
Sl4CL02 | 699 | 77723.64 | 5.64 | 37.7 | -0.126 | 10926 | Peroxisome |
Sl4CL03 | 560 | 61700.2 | 5.49 | 33.69 | -0.028 | 8735 | Peroxisome |
Sl4CL04 | 957 | 102255.02 | 5.45 | 31.47 | 0.476 | 14550 | Peroxisome |
Sl4CL05 | 538 | 58930.46 | 8.49 | 30.69 | 0.07 | 8369 | Peroxisome |
Sl4CL06 | 548 | 60394.02 | 5.9 | 32.36 | 0.045 | 8590 | Peroxisome |
Table 2 Physico-chemical properties of 4CL family proteins in tomato(Solanum lycopersicum)
基因名称Gene name | 氨基酸个数 Number of amino acids | 分子量 Molecular weight/kD | 等电点 Isoelectric point/pI | 不稳定指数Instability index | 总平均亲水系数Grand average of hydropathicity | 原子总数Total number of atoms | 亚细胞定位Subcellular localization |
---|---|---|---|---|---|---|---|
Sl4CL01 | 570 | 62080.54 | 5.34 | 34.92 | 0.106 | 8820 | Peroxisome Chloroplast |
Sl4CL02 | 699 | 77723.64 | 5.64 | 37.7 | -0.126 | 10926 | Peroxisome |
Sl4CL03 | 560 | 61700.2 | 5.49 | 33.69 | -0.028 | 8735 | Peroxisome |
Sl4CL04 | 957 | 102255.02 | 5.45 | 31.47 | 0.476 | 14550 | Peroxisome |
Sl4CL05 | 538 | 58930.46 | 8.49 | 30.69 | 0.07 | 8369 | Peroxisome |
Sl4CL06 | 548 | 60394.02 | 5.9 | 32.36 | 0.045 | 8590 | Peroxisome |
基因名称 Gene name | α-螺旋α-helix/% | 延伸链 Extended strand/% | β-转角 β-turn/% | 无规则卷曲 Random coil/% |
---|---|---|---|---|
Sl4CL01 | 32.28 | 18.6 | 6.84 | 42.28 |
Sl4CL02 | 33.19 | 18.6 | 9.01 | 39.2 |
Sl4CL03 | 30.89 | 20.71 | 7.5 | 40.89 |
Sl4CL04 | 41.69 | 15.57 | 7.94 | 34.8 |
Sl4CL05 | 30.67 | 20.07 | 8.36 | 40.89 |
Sl4CL06 | 31.93 | 19.53 | 8.58 | 39.96 |
Table 3 Secondary structure of tomato 4CL gene family
基因名称 Gene name | α-螺旋α-helix/% | 延伸链 Extended strand/% | β-转角 β-turn/% | 无规则卷曲 Random coil/% |
---|---|---|---|---|
Sl4CL01 | 32.28 | 18.6 | 6.84 | 42.28 |
Sl4CL02 | 33.19 | 18.6 | 9.01 | 39.2 |
Sl4CL03 | 30.89 | 20.71 | 7.5 | 40.89 |
Sl4CL04 | 41.69 | 15.57 | 7.94 | 34.8 |
Sl4CL05 | 30.67 | 20.07 | 8.36 | 40.89 |
Sl4CL06 | 31.93 | 19.53 | 8.58 | 39.96 |
Fig.6 Expression patterns of tomato 4CL genes under different concentrations of nitrogen Histograms with different letters indicate significant difference at the 0.05 level
Fig.8 Subcellular localization of Sl4CL03 in Nicotiana tabacum leaf cells A-D:pCAMBIA1302 empty vector fluorescence,chloroplast fluorescence,bright,and merged.E-H:Sl4CL03 protein fluorescence,chloroplast fluorescence,bright,and merged
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