Ma P, Sivaloganathan B, Reddy P K, Chan W K, Lam T J
Department of Biological Sciences, National University of Singapore, 10 Kent Ridge Crescent, Singapore 119260.
Mar Biotechnol (NY). 2001 Sep;3(5):463-9. doi: 10.1007/s10126-001-0018-1.
The enzymatic activities of alpha-amylase and its corresponding messenger RNA levels in developing sea bass (Lates calcarifer) larvae were studied from hatching until 27 days post hatching (dph). An increasing activity of amylase enzyme was measured until 5 dph, and the activity gradually decreased thereafter and reached a constant level by 12 dph. To achieve a better understanding of the molecular mechanisms underlying amylase expression, we have cloned and sequenced a 318-bp fragment of alpha-amylase complementary DNA. Based on this sequence, a real-time reverse transcriptase polymerase chain reaction technique to monitor the changes in the mRNA levels in the larvae was developed. A correlation between enzymatic activity and mRNA level of alpha-amylase could be demonstrated during the early development of sea bass larvae. This suggests that the changes in alpha-amylase are controlled at least at the transcriptional level during early larval development of sea bass.
研究了尖吻鲈(Lates calcarifer)幼体从孵化到孵化后27天(dph)期间α-淀粉酶的酶活性及其相应信使RNA水平。在5 dph之前测定到淀粉酶活性不断增加,此后活性逐渐下降,到12 dph时达到稳定水平。为了更好地理解淀粉酶表达的分子机制,我们克隆并测序了α-淀粉酶互补DNA的一个318 bp片段。基于该序列,开发了一种实时逆转录聚合酶链反应技术来监测幼体中mRNA水平的变化。在尖吻鲈幼体的早期发育过程中,可以证明α-淀粉酶的酶活性与mRNA水平之间存在相关性。这表明在尖吻鲈幼体早期发育过程中,α-淀粉酶的变化至少在转录水平上受到控制。