摘要
提取鹰爪虾眼柄总RNA,以眼柄总RNA为模板,根据日本对虾的具有蜕皮抑制活性的神经肽Pej SGP Ⅳ的氨基酸设计兼并引物,进行RT PCR扩增,在适宜条件下得到一特异性片段。将这一特异性片段克隆到载体中进行测序,测得鹰爪虾的特异性cDNA片段由213个碱基对组成,推测的氨基酸序列由71个氨基酸组成。序列比较发现许多甲壳动物的CHH家族神经肽与它相似。在所有与该序列推测的氨基酸序列相似的甲壳动物CHH家族神经肽中,MIH与它的相似度最高,表明由鹰爪虾眼柄特异性cDNA片段推定的氨基酸序列可能是鹰爪虾的MIH片段,它相当于MIH成熟肽的第5至75个氨基酸。
Total RNAs were extracted from eyestalks of the shrimp Trachypenaeus curvirostris. Eyestalk complementary DNAs were obtained from these total RNAs by reverse transcription. Based on the amino acid sequence of the neuropeptide PejSGPⅣ which has moltinhibiting activity from Marsupenaeus japonicus, two degenerate primers were designed. Polymerase Chain Reaction (PCR) was performed using these primers and these eyestalk cDNAs. A specific cDNA was amplified under proper conditions. The specific cDNA was cloned into vector for sequencing. The sequencing results demonstrated that the specific cDNA from the shrimp T. curvirostris consisted of 213 base pairs. The alignment of the deduced amino acid sequence from eyestalk cDNA of the shrimp T. curvirostris with neuropeptides of CHH family from various crustaceans demonstrated that it had the highest identity and homology with most MIHs from various crustaceans. It suggested that the deduced amino acid sequence from eyestalk cDNA of the shrimp T. curvirostris was a part of MIH neuropeptide of T. curvirostris. The sequence corresponded to the amino acid sequence from 5 to 75 residues of MIH.
出处
《水产学报》
CAS
CSCD
北大核心
2002年第6期487-492,共6页
Journal of Fisheries of China
基金
国家自然科学基金资助项目(40106018)
中国科学院知识创新资助项目(ZKCX2 211)
关键词
鹰爪虾
蜕皮抑制激素
CDNA克隆
序列分析
基因克隆
神经肽
Trachypenaeus curvirostris
molt-inhibiting hormone
complementary DNA cloning
sequence analysis
