An Efficient Protocol for DNA Amplification of Multiple Amphibian Skin Antimicrobial Peptide cDNAs
An Efficient Protocol for DNA Amplification of Multiple Amphibian Skin Antimicrobial Peptide cDNAs
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DOI：
10.1007/978-1-60761-535-4_13
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发表时间：
2010-01-01
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影响因子：
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通讯作者：
Kobayashi, Tetsuya
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文献类型：
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作者：
Iwamuro, Shawichi;Kobayashi, Tetsuya
抗菌肽（AMPs）在许多生物体的宿主先天防御系统中发挥重要作用。两栖动物的皮肤有望成为新型抗菌肽的特别丰富的来源。在两栖动物中，amp是由前体蛋白通过加工酶的特定切割产生的。虽然前体中AMP编码区的核苷酸序列是高可变的，但其他区域，包括5'-和3'-非翻译区（utr）的核苷酸序列在不同的前体中是高度或相对保守的。这种核苷酸序列的保守性为利用cDNA末端的X-rapid扩增（T-RACE）和逆转录聚合酶链反应（RT-PCR）方法克隆抗菌肽基因提供了一种有效的策略。考虑到这一策略，我们已经建立了一种有效的方案，适用于扩增编码两栖动物AMP前体蛋白的多种cdna。
Antimicrobial peptides (AMPs) play an important role in the host's innate defence system in many organisms. Amphibian skin is expected to be a particularly rich Source of novel AMPs. In amphibians, AMPs are produced from precursor proteins via specific cleavage by processing enzymes. While the nucleotide sequences of the AMP coding region in precursors are hypervariable, those of other regions, including the 5'- and 3'-untranslated regions (UTRs), are highly or relatively conserved in different precursors. Such nucleotide sequence conservation Suggests an efficient strategy for Molecular cloning of the antimicrobial peptide genes by X-rapid amplification of cDNA ends (T-RACE) and reverse transcriptase polymerase chain reaction (RT-PCR) methods using specific primers. With this strategy in mind we have established an efficient protocol suitable for amplification of multiple cDNAs encoding amphibian AMP precursor proteins.