Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

doi:10.3969/j.issn.1004-1524.20230104

Acta Agriculturae Zhejiangensis ›› 2024, Vol. 36 ›› Issue (2): 247-253.DOI: 10.3969/j.issn.1004-1524.20230104

• Animal Science • Previous Articles Next Articles

Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

PENG Jiacheng¹(), WU Yue¹, XU Jiehao², XIA Meiwen¹, QI Tianpeng¹, XU Haisheng¹^,³^,^*()

1. College of Animal Sciences, Zhejiang University, Hangzhou 310058, China
2. College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, Zhejiang, China
3. Zhejiang University Huzhou South Taihu Lake Modern Agricultural Science and Technology Promotion Center, Huzhou 313000, Zhejiang, China

Received:2023-02-06 Online:2024-02-25 Published:2024-03-05

Abstract

Abstract:

Cadmium (Cd), with strong accumulation and toxicity, is one of the non-essential heavy metal elements, which could cause great harm to aquatic organisms. To better understand the molecular toxicity of cadmium to Macrobrachium nipponense, and provide biomarkers for ecological risk warning and assessment of cadmium pollution in water environment. In this paper, paxillin gene (MnPXN) of M. nipponense was cloned, and the effect of cadmium stress on its expression was examined. The total length of MnPXN gene is 3 655 bp, including 1 491 bp coding region, encoding 496 amino acids; 6 low complexity regions and 3 conservative LIM domains are included in MnPXN, forming a branch with Litopenaeus vannamei. It is closely related to insects such as silkworm and Tribolium castaneum, and is farthest related to nematodes. The results of quantitative reverse transcriptase-mediated PCR (qRT-PCR) showed that the expression level of MnPXN was high in hepatopancreas, stomach and gill, but low in heart and muscle. The expression level of MnPXN gene in hepatopancreas that after 24 h of treatment with 0.242 7 mg·L^-1 CdCl₂ was significantly higher than that in control group; After 6 h of 0.020 0 mg·L^-1 CdCl₂ stress, the expression level of MnPXN gene increased significantly, reaching the maximum at 24 h, and gradually recovered to the initial level at 96 h. Besides, RNA interference could significantly reduce the expression level of MnPXN in hepatopancreas and significantly increase the mortality of M. nipponense, indicating that MnPXN could reduce the toxic effect of cadmium stress on M. nipponense, and its mechanism needs further study.

Key words: cadmium stress, Macrobrachium nipponense, paxillin, gene expression, RNA interference

CLC Number:

S917.4

PENG Jiacheng, WU Yue, XU Jiehao, XIA Meiwen, QI Tianpeng, XU Haisheng. Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression[J]. Acta Agriculturae Zhejiangensis, 2024, 36(2): 247-253.

Figures/Tables 6

References 20

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引物 Primer	引物序列 Primer sequence(5'→3')	用途 Purpose
MnPXN-3'-F₁	CAATGCCAGCAGCCTCCAGACAC	RACE
MnPXN-3'-R₁	AAGCCGTCCTCGCCGAAGGTGTT	RACE
MnPXN-3'-F₂	TGGACGAGTTGAACAGCGTTGACA	RACE
MnPXN-3'-R₂	TCCCTTCTGGGTCGTCGTGATGC	RACE
MnPXN-3'-F₃	GAATGACCTGAGTGGCACCTGAC	RACE
MnPXN-3'-R₃	TGCCGTGAAGAATGACATAGAAT	RACE
MnPXN-3'-F₄	GAGGAAGTGCGGACGGAGCAGAGGT	RACE
MnPXN-3'-R₄	GATCTCGATGTGCCGTGAAGAAT	RACE
MnPXN-3'-F₅	GTTATGTCATTGGCTCAAGAGGA	RACE
MnPXN-3'-R₅	GTGTATTTATACTGTATGCCTAACC	RACE
MnPXN-3'-F₆	GCTACAAGAGGCTCCCAACTACAT	RACE
MnPXN-3'-R₆	CAGGAGGTAAAGACTACTGATGT	RACE
MnPXN-3'-F₇	AAAAGGTTTGGTAAATGTCCAGGTT	RACE
MnPXN-3'-R_UPM	CTAATACGACTCACTATAGGGC	RACE
MnPXN-5'-F₁	TTGTTCATCCAGGAGGTAAAGAC	RACE
MnPXN-5'-R₁	GCCAATAAGGCATCAATGGTATCCTG	RACE
MnPXN-5'-F₂	GAAGTTTGGCAGTATCTTATGGA	RACE
MnPXN-5'-R₂	TTGTGAATGAGGGTGGCGTTTGG	RACE
MnPXN-5'-F₃	CAATAAGGCATCAATGGTATCCTGCA	RACE
MnPXN-5'-R₃	AATTTCAAAAATCTAATT	RACE
MnPXN-5'-F_UPM	CTAATACGACTCACTATAGGGCAA GCAGTGGTATCAACGCAGAGT	RACE
MnPXN-5'-R₄	GGGCATCATTCTTTGTCTTTCTT	RACE
MnPXN-RT-F	CCCCATCTTGGATAAATGCGTGAC	qRT-PCR
MnPXN-RT-R	AAGCCGTCCTCGCCGAAGGTGTTGC	qRT-PCR
β-actin-F	GTGCCCATCTACGAGGGTTA	qRT-PCR
β-actin-R	CGTCAGGGAGCTCGTAAGAC	qRT-PCR

引物 Primer	引物序列 Primer sequence(5'→3')	用途 Purpose
MnPXN-3'-F₁	CAATGCCAGCAGCCTCCAGACAC	RACE
MnPXN-3'-R₁	AAGCCGTCCTCGCCGAAGGTGTT	RACE
MnPXN-3'-F₂	TGGACGAGTTGAACAGCGTTGACA	RACE
MnPXN-3'-R₂	TCCCTTCTGGGTCGTCGTGATGC	RACE
MnPXN-3'-F₃	GAATGACCTGAGTGGCACCTGAC	RACE
MnPXN-3'-R₃	TGCCGTGAAGAATGACATAGAAT	RACE
MnPXN-3'-F₄	GAGGAAGTGCGGACGGAGCAGAGGT	RACE
MnPXN-3'-R₄	GATCTCGATGTGCCGTGAAGAAT	RACE
MnPXN-3'-F₅	GTTATGTCATTGGCTCAAGAGGA	RACE
MnPXN-3'-R₅	GTGTATTTATACTGTATGCCTAACC	RACE
MnPXN-3'-F₆	GCTACAAGAGGCTCCCAACTACAT	RACE
MnPXN-3'-R₆	CAGGAGGTAAAGACTACTGATGT	RACE
MnPXN-3'-F₇	AAAAGGTTTGGTAAATGTCCAGGTT	RACE
MnPXN-3'-R_UPM	CTAATACGACTCACTATAGGGC	RACE
MnPXN-5'-F₁	TTGTTCATCCAGGAGGTAAAGAC	RACE
MnPXN-5'-R₁	GCCAATAAGGCATCAATGGTATCCTG	RACE
MnPXN-5'-F₂	GAAGTTTGGCAGTATCTTATGGA	RACE
MnPXN-5'-R₂	TTGTGAATGAGGGTGGCGTTTGG	RACE
MnPXN-5'-F₃	CAATAAGGCATCAATGGTATCCTGCA	RACE
MnPXN-5'-R₃	AATTTCAAAAATCTAATT	RACE
MnPXN-5'-F_UPM	CTAATACGACTCACTATAGGGCAA GCAGTGGTATCAACGCAGAGT	RACE
MnPXN-5'-R₄	GGGCATCATTCTTTGTCTTTCTT	RACE
MnPXN-RT-F	CCCCATCTTGGATAAATGCGTGAC	qRT-PCR
MnPXN-RT-R	AAGCCGTCCTCGCCGAAGGTGTTGC	qRT-PCR
β-actin-F	GTGCCCATCTACGAGGGTTA	qRT-PCR
β-actin-R	CGTCAGGGAGCTCGTAAGAC	qRT-PCR

Cloning of paxillin gene from Macrobrachium nipponense and effect of cadmium stress on its expression

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