利用cDNA文库构建分析伊朗蝎毒腺中一些推测的新肽

IF 1 Q4 PHARMACOLOGY & PHARMACY Jundishapur Journal of Natural Pharmaceutical Products Pub Date : 2023-02-04 DOI:10.5812/jjnpp-133423
A. Jalali, M. Baradaran, H. Galehdari, Parnian Ghoncheh, Maryam Naderi Soorki
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引用次数: 0

摘要

背景:迄今为止,在蝎子毒液中已经鉴定出超过30万种不同的肽。然而,只有有限数量的这些肽被描述。目的:对伊朗黄蝎“半蝎”毒液腺cDNA文库中一些推测的多肽进行了鉴定。方法:提取黄蝎腺体总RNA。采用聚合酶链式反应(PCR)合成单链cDNA (sscDNA)和双链cDNA (dscDNA)。通过将dscDNA插入到一个特殊的载体中,然后将其转化为具有化学活性的大肠杆菌作为宿主,获得了cDNA文库。通过在溶根菌肉汤(LB)-琼脂平板上培养液体文库来筛选文库。对阳性克隆进行质粒提取和插入片段测序分析。最后,利用生物信息学软件对所有cDNA序列进行分析和表征。结果:随机分析100个菌落。89条cDNA序列质量可用于生物信息学分析。选取5个序列进行进一步分析。将与这些序列相关的肽分为非二硫桥肽(NDBP)和二硫桥肽(DBP)两类,并提出了它们在健康和医疗问题上的应用。结论:本研究获得的数据可能为进一步的体内和体外功能分析提供重要的资源,以鉴定有价值的治疗肽。
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Analysis of Some Putative Novel Peptides from Iranian Scorpion Venom Glands, Hemiscorpius lepturus, Using cDNA Library Construction
Background: To date, more than 300,000 distinct peptides have been identified in scorpion venom. However, only a limited number of these peptides have been described. Objectives: We characterized some putative peptides from the venom gland cDNA library of the Iranian yellow scorpion Hemiscorpius lepturus”. Methods: Total RNA was extracted from yellow Iranian scorpion glands. Single-stranded cDNA (sscDNA) and double-stranded cDNA (dscDNA) were synthesized by polymerase chain reaction (PCR). A cDNA library was achieved by inserting dscDNA into a special vector and subsequently transformed to chemically competent Escherichia coli as a host. The library was screened by culturing the liquid library on Lysogeny broth (LB)-agar plates. Analysis of positive clones was performed by plasmid extraction and the sequencing of the inserts. Finally, all cDNA sequences were analyzed and characterized by bioinformatics software. Results: One hundred colonies were randomly analyzed. Eighty-nine cDNA sequences had acceptable quality for bioinformatics analysis. Five sequences were selected for further analysis. The peptides related to these sequences were divided into two groups, non-disulfide bridge peptides (NDBP) and disulfide bridge peptides (DBP), the application of which in health and medical issues has been suggested. Conclusions: The data obtained in this study may be an important resource for further in vivo and in vitro functional assays to identify valuable therapeutic peptides.
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