Molecular cloning and characterisation of the PmEglin cDNA in the leech Hirudinaria sp.

Cheng, Biao; Shao, Gui-Yan; Li, Yuan; Tian, Qing-Qing; Wang, Suyan; Liu, Fei

doi:10.1007/s11756-022-01277-3

Cited by 2 publications

(3 citation statements)

References 78 publications

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“…The corresponding protein (PmEglin) of TRINITY_DN808_c0_g1 belongs to the potato type I proteinase inhibitor family, which can alter physiological processes by regulating immune responses. Their mRNA expression trends are both consistent with the data in external transcriptome databases (Cheng et al, 2022;Shao et al, 2022).…”

Section: Discussionsupporting

confidence: 84%

“…The expression of this kind of genes is induced by blood meal ingestion. When the protein storage of these genes in salivary glands is su cient, their mRNA expression will decrease gradually to their pre-feeding levels (Cheng et al 2019;Cheng et al, 2022). The expression trend of HspHirudin mRNA during 3 feeding periods (H0, Hing and H1) was consistent with the trend of transcript TRINITY_DN14935_c0_g2 in the external transcriptome database.…”

Section: Discussionsupporting

confidence: 54%

“…In this study, the other two of the 10 unigenes we screened were unigenes of immunolog-related proteins, eglin C (TRINITY_DN808_c0_g1) and C-type lectins (TRINITY_DN11316_c0_g1). They were found in leeches or other haemophiles, and were associated with blood meal ingestion (Bode et al,1986;Seemüller et al, 1977;Das et al, 2010;Polanska et al, 2020;Cheng et al, 2022). We found out that a variety of unigenes with functions related to inhibiting blood clotting and immunomodulatory were upregulated in the salivary gland of Hirudinaria sp.…”

Section: Discussionmentioning

confidence: 62%

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Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

Cheng,

Hu,

Wang

et al. 2024

Biologia

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BackgroundHirudinaria sp., as one of the origins of leeches used in traditional Chinese medicine, is proved clinically effective in treating cardiovascular and cerebrovascular diseases. Substances with pharmacological activity are stored in salivary glands of leeches. Among these substances, hirudin exhibits high anti-thrombin activity. However, the structure, function and expression pattern of HspHirudin in Hirudinaria sp. distributed in Guangxi are still unclear. Methods and resultsIllumina HiSeq platform was used to obtain the transcriptome datasets of salivary glands from Hirudinaria sp. during different feeding periods. Unigenes which were signi cantly up-regulated after a blood meal and had functions related to the processes of blood meal ingestion were identi ed. The function of HspHirudin was studied based on gene cloning, bioinformatics analysis, prokaryotic expression analysis and anti-thrombin activity analysis, and its expression pattern was characterized by quantitative realtime PCR. The reads were assembled into 167564 unigenes, and ten genes with functions related to the processes of blood meal ingestion in salivary glands of Hirudinaria sp. were identi ed. After alignment, transcript (TRINITY_DN14935_c0_g2) was annotated to Hirudin-P6 (2E-31) from Poecilobdella manillensis and named HspHirudin. The HspHirudin cDNA was 249 bp in length, and it encodes a mature peptide of 82 aa and a signal peptide of 20 aa. The mature HspHirudin presented typical structural characteristics of hirudins: 3 pairs of disul de bonds, the PKP and DFxxIP motifs. HspHirudin and Hirudin-P6 shared similar structure of N-terminus 3 aa and C-terminus, and their main differences were observed in the spherical structure domain. By constructing a prokaryotic expression system, HspHirudin was proven to have anti-thrombin activity. The high expression of HspHirudin mRNA was induced by bloodmeal feeding. ConclusionsTen genes with functions related to the processes of blood meal ingestion were screened from transcriptome datasets of salivary glands from Hirudinaria sp. during different feeding periods. Among them, HspHirudin, which belongs to the family of hirudins, has anti-thrombin activity. The results of the study provide theoretical basis for further development and utilization of Hirudinaria sp.. HighlightsTranscripts from transcriptome datasets of salivary glands from Hirudinaria during different feeding periods were screened.HspHirudin, which belongs to the family of hirudins, has anti-thrombin activity.The high expression of HspHirudin mRNA was induced by bloodmeal feeding.

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Section: Discussionsupporting

confidence: 84%

Section: Discussionsupporting

confidence: 54%

Section: Discussionmentioning

confidence: 62%

See 1 more Smart Citation

Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

Cheng,

Hu,

Wang

et al. 2024

Biologia

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Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

Cheng

Wang

et al. 2023

Preprint

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Background Hirudinaria sp., as one of the origins of leeches used in traditional Chinese medicine, is proved clinically effective in treating cardiovascular and cerebrovascular diseases. Substances with pharmacological activity are stored in salivary glands of leeches. Among these substances, hirudin exhibits high anti-thrombin activity. However, the structure, function and expression pattern of HspHirudin in Hirudinaria sp. distributed in Guangxi are still unclear.Methods and results Illumina HiSeq platform was used to obtain the transcriptome datasets of salivary glands from Hirudinaria sp. during different feeding periods. Unigenes which were significantly up-regulated after a blood meal and had functions related to the processes of blood meal ingestion were identified. The function of HspHirudin was studied based on gene cloning, bioinformatics analysis, prokaryotic expression analysis and anti-thrombin activity analysis, and its expression pattern was characterized by quantitative real-time PCR. The reads were assembled into 167564 unigenes, and ten genes with functions related to the processes of blood meal ingestion in salivary glands of Hirudinaria sp. were identified. After alignment, transcript (TRINITY_DN14935_c0_g2) was annotated to Hirudin-P6 (2E-31) from Poecilobdella manillensis and named HspHirudin. The HspHirudin cDNA was 249 bp in length, and it encodes a mature peptide of 82 aa and a signal peptide of 20 aa. The mature HspHirudin presented typical structural characteristics of hirudins: 3 pairs of disulfide bonds, the PKP and DFxxIP motifs. HspHirudin and Hirudin-P6 shared similar structure of N-terminus 3 aa and C-terminus, and their main differences were observed in the spherical structure domain. By constructing a prokaryotic expression system, HspHirudin was proven to have anti-thrombin activity. The high expression of HspHirudin mRNA was induced by bloodmeal feeding.Conclusions Ten genes with functions related to the processes of blood meal ingestion were screened from transcriptome datasets of salivary glands from Hirudinaria sp. during different feeding periods. Among them, HspHirudin, which belongs to the family of hirudins, has anti-thrombin activity. The results of the study provide theoretical basis for further development and utilization of Hirudinaria sp..

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Molecular cloning and characterisation of the PmEglin cDNA in the leech Hirudinaria sp.

Cited by 2 publications

References 78 publications

Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

Salivary gland transcriptome analysis in Hirudinaria sp. during different feeding periods and functional studies of HspHirudin

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