浙江农业学报 ›› 2022, Vol. 34 ›› Issue (4): 695-705.DOI: 10.3969/j.issn.1004-1524.2022.04.05
余艳玲1(), 罗洪林1, 罗辉2, 冯鹏霏1, 潘传燕1, 宋漫玲1, 肖蕊1, 张永德1,*(
)
收稿日期:
2020-10-13
出版日期:
2022-04-25
发布日期:
2022-04-28
通讯作者:
张永德
作者简介:
*张永德,E-mail: yondar@126.com基金资助:
YU Yanling1(), LUO Honglin1, LUO Hui2, FENG Pengfei1, PAN Chuanyan1, SONG Manling1, XIAO Rui1, ZHANG Yongde1,*(
)
Received:
2020-10-13
Online:
2022-04-25
Published:
2022-04-28
Contact:
ZHANG Yongde
摘要:
为研究卵形鲳鲹生肌调节因子(MRF)在胚胎发育中所发挥的重要作用,对卵形鲳鲹MRF基因家族进行了全基因组鉴定和生物信息学分析,并对其在13个胚胎发育阶段的基因表达进行了定量分析。结果表明:在卵形鲳鲹基因组中共鉴定出5个MRF基因家族成员:MyoD1、MyoD2、Myf5、Myf6和MyoG,分别编码297、263、240、231、250个氨基酸。MRF家族基因存在典型的BASIC与HLH结构域,其中MyoD1、MyoD2与MyoG分别定位于9号、1号与2号染色体,而Myf5与Myf6基因均定位于22号染色体且位于同一基因座。MRF家族基因胚胎定量表达研究表明,MyoG在受精卵到胚体形成期表达量极低,在眼囊、耳囊与心脏跳动期表达量迅速升高,而到晶体出现期表达量又有所降低,推测其可能在眼囊、耳囊与心脏跳动期发挥主要的生肌调节作用,而后期在维持肌肉形态或肌肉发育中仍发挥重要作用。MyoD1、MyoD2、Myf5与Myf6在受精卵到原肠中期表达量较低,而从原肠末期到晶体出现期表达量迅速升高(P<0.05),表明其主要从原肠末期开始发挥生肌调控作用。结果表明,MRF家族成员在胚胎发育过程中发挥重要的作用,但不同的MRF家族成员发挥作用的时间及功能可能不同。
中图分类号:
余艳玲, 罗洪林, 罗辉, 冯鹏霏, 潘传燕, 宋漫玲, 肖蕊, 张永德. 卵形鲳鲹生肌调节因子基因家族的鉴定及在胚胎中的表达[J]. 浙江农业学报, 2022, 34(4): 695-705.
YU Yanling, LUO Honglin, LUO Hui, FENG Pengfei, PAN Chuanyan, SONG Manling, XIAO Rui, ZHANG Yongde. Genome-wide identification and expression of MRF gene family in embryonic development of Trachinotus ovatus[J]. Acta Agriculturae Zhejiangensis, 2022, 34(4): 695-705.
引物名称Primer | 上游引物Forward primer (5'-3') | 下游引物Reverse primer (5'-3') |
---|---|---|
MyoD1 | AACGCCATCAGCTACATCGAG | ACAGGTGGGGCCGTTAAAATC |
MyoD2 | CGGCCTACTGAAGCCCGAA | CTCATGGTTGCTGCTTTTCTCC |
Myf5 | AAGAACGAGAGTTTGGGCGA | AGGACGTGGTATATGGGCCT |
Myf6 | TCAGCTACATCGAGCGGTT | TCCAGTGGTACTCATGACCG |
MyoG | CTTCTACGAGGGAGGGGACA | GCTGGATGGAGAGGCTTTGT |
18S rRNA | GACTCGGGGAGGTAGTGACG | AGATACGCTATTGGAGCTGGAA |
表1 卵形鲳鲹MRF家族基因qRT-PCR引物信息
Table 1 qRT-PCR primers used for T. ovatus MRF family genes
引物名称Primer | 上游引物Forward primer (5'-3') | 下游引物Reverse primer (5'-3') |
---|---|---|
MyoD1 | AACGCCATCAGCTACATCGAG | ACAGGTGGGGCCGTTAAAATC |
MyoD2 | CGGCCTACTGAAGCCCGAA | CTCATGGTTGCTGCTTTTCTCC |
Myf5 | AAGAACGAGAGTTTGGGCGA | AGGACGTGGTATATGGGCCT |
Myf6 | TCAGCTACATCGAGCGGTT | TCCAGTGGTACTCATGACCG |
MyoG | CTTCTACGAGGGAGGGGACA | GCTGGATGGAGAGGCTTTGT |
18S rRNA | GACTCGGGGAGGTAGTGACG | AGATACGCTATTGGAGCTGGAA |
蛋白质名称 Protein name | 蛋白质编号 Locus ID | 蛋白质长度 Size/aa | 相对分子质量 Molecular weight/u | 等电点 Pi | 不稳定指数 Instability index | 疏水性均值 GRAVY |
---|---|---|---|---|---|---|
ToMyoD1 | EVM0022498 | 297 | 32 206.38 | 5.33 | 71.51 | -0.591 |
ToMyoD2 | EVM0018159 | 263 | 28 731.83 | 5.87 | 66.57 | -0.634 |
ToMyf5 | EVM0020882 | 240 | 26 187.14 | 6.23 | 89.12 | -0.594 |
ToMyf6 | EVM0005068 | 231 | 25 542.27 | 5.93 | 65.35 | -0.849 |
ToMyoG | EVM0008148 | 250 | 27 421.64 | 6.33 | 65.97 | -0.650 |
表2 卵形鲳鲹MRF家族蛋白质的理化特性
Table 2 Physicochemical properties of MRF family proteins in T. ovatus
蛋白质名称 Protein name | 蛋白质编号 Locus ID | 蛋白质长度 Size/aa | 相对分子质量 Molecular weight/u | 等电点 Pi | 不稳定指数 Instability index | 疏水性均值 GRAVY |
---|---|---|---|---|---|---|
ToMyoD1 | EVM0022498 | 297 | 32 206.38 | 5.33 | 71.51 | -0.591 |
ToMyoD2 | EVM0018159 | 263 | 28 731.83 | 5.87 | 66.57 | -0.634 |
ToMyf5 | EVM0020882 | 240 | 26 187.14 | 6.23 | 89.12 | -0.594 |
ToMyf6 | EVM0005068 | 231 | 25 542.27 | 5.93 | 65.35 | -0.849 |
ToMyoG | EVM0008148 | 250 | 27 421.64 | 6.33 | 65.97 | -0.650 |
图3 卵形鲳鲹MRF基因家族的结构域及保守基序 a,MRF家族成员的结构域及保守基序位置;b,MRF家族成员的保守基序组成。
Fig.3 Domain and conserved motif of MRF gene family in T.ovatus a, Domain and conserved motif position of MRF family genes; b, Conserved motif composition of MRF family genes.
物种 Species | 分类 Taxonomy | 基因名称 Gene name | 蛋白质编号 Locus ID | 蛋白质长度 Size/aa |
---|---|---|---|---|
高体鰤 | 鲹形目Carangiformes; | SdMyoD1 | ENSSDUP00000023227 | 298 |
S. dumerili | 鲹科Carangidae; | SdMyoD2 | ENSSDUP00000025164 | 262 |
鰤属Seriola | SdMyf5 | ENSSDUP00000025182 | 240 | |
SdMyf6 | ENSSDUP00000025208 | 238 | ||
SdMyoG | ENSSDUP00000002433 | 250 | ||
黄尾鰤 | 鲹形目Carangiformes; | SlMyoD1 | ENSSLDP00000028917 | 298 |
S. lalandi dorsalis | 鲹科Carangidae; | SlMyoD2 | ENSSLDP00000027557 | 263 |
鰤属Seriola | SlMyf5 | ENSSLDP00000024335 | 240 | |
SlMyf6 | ENSSLDP00000024364 | 239 | ||
SlMyoG | ENSSLDP00000009264 | 250 | ||
半滑舌鳎 | 鲽形目Pleuronectiformes; | CsMyoD1 | ENSCSEP00000003405 | 296 |
C. semilaevis | 舌鳎科Cynoglossidae; | CsMyoD2 | ENSCSEP00000016973 | 214 |
舌鳎属Cynoglossus | CsMyf5 | ENSCSEP00000006862 | 246 | |
CsMyf6 | ENSCSEP00000006878 | 241 | ||
CsMyoG | ENSCSEP00000007229 | 243 | ||
大菱鲆 | 鲽形目Pleuronectiformes; | SmMyoD1 | ENSSMAP00000005978 | 297 |
S. maximus | 菱鲆科Scophthalmidae; | SmMyoD2 | ENSSMAP00000010597 | 265 |
菱鲆属 Scophthalmus | SmMyf5 | ENSSMAP00000007369 | 297 | |
SmMyf6 | ENSSMAP00000007390 | 269 | ||
SmMyoG | ENSSMAP00000010312 | 246 |
表3 四种鱼类MRF家族基因信息
Table 3 Information of the MRF family genes in four fish species
物种 Species | 分类 Taxonomy | 基因名称 Gene name | 蛋白质编号 Locus ID | 蛋白质长度 Size/aa |
---|---|---|---|---|
高体鰤 | 鲹形目Carangiformes; | SdMyoD1 | ENSSDUP00000023227 | 298 |
S. dumerili | 鲹科Carangidae; | SdMyoD2 | ENSSDUP00000025164 | 262 |
鰤属Seriola | SdMyf5 | ENSSDUP00000025182 | 240 | |
SdMyf6 | ENSSDUP00000025208 | 238 | ||
SdMyoG | ENSSDUP00000002433 | 250 | ||
黄尾鰤 | 鲹形目Carangiformes; | SlMyoD1 | ENSSLDP00000028917 | 298 |
S. lalandi dorsalis | 鲹科Carangidae; | SlMyoD2 | ENSSLDP00000027557 | 263 |
鰤属Seriola | SlMyf5 | ENSSLDP00000024335 | 240 | |
SlMyf6 | ENSSLDP00000024364 | 239 | ||
SlMyoG | ENSSLDP00000009264 | 250 | ||
半滑舌鳎 | 鲽形目Pleuronectiformes; | CsMyoD1 | ENSCSEP00000003405 | 296 |
C. semilaevis | 舌鳎科Cynoglossidae; | CsMyoD2 | ENSCSEP00000016973 | 214 |
舌鳎属Cynoglossus | CsMyf5 | ENSCSEP00000006862 | 246 | |
CsMyf6 | ENSCSEP00000006878 | 241 | ||
CsMyoG | ENSCSEP00000007229 | 243 | ||
大菱鲆 | 鲽形目Pleuronectiformes; | SmMyoD1 | ENSSMAP00000005978 | 297 |
S. maximus | 菱鲆科Scophthalmidae; | SmMyoD2 | ENSSMAP00000010597 | 265 |
菱鲆属 Scophthalmus | SmMyf5 | ENSSMAP00000007369 | 297 | |
SmMyf6 | ENSSMAP00000007390 | 269 | ||
SmMyoG | ENSSMAP00000010312 | 246 |
图7 MRF家族基因在卵形鲳鲹不同胚胎发育阶段的表达 没有相同小写字母表示差异显著(P<0.05)。
Fig.7 Expression pattern of MRF family genes in different developmental stages of T. ovatus embryos The bars without the same letters meant the significant difference (P<0.05).
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