Vasa, which is a conserved member of the DEAD-box protein family, plays an indispensable role in primordial germ cell proliferation. However, the expression ofvasa gene during the reproductive cycle in ovoviviparous f...Vasa, which is a conserved member of the DEAD-box protein family, plays an indispensable role in primordial germ cell proliferation. However, the expression ofvasa gene during the reproductive cycle in ovoviviparous fish has not been documented. In this study, the full-length sequence of vasa was obtained from the ovary of Korean rockfish （Sebastes schlegeli） using reverse transcription-PCR and rapid amplification of cDNA ends. The Vasa with a mature protein of 650 amino acids showed greatest homology （84%） with giant gourami （Osphronemus gorarny） and Pacific bluefin tuna （Thunnus orientalis）. The expression of vasa mRNA in Korean rockfish was detected in gonads only, suggesting its specific role in gonadal development. In addition, seasonal changes in the vasa expression levels were examined in gonads by quantitative real-time PCR. The vasa transcript levels in adult testis were found higher during spermatogenesis than during spermiation. The vasa transcript levels remained relatively high at the early ovary stage but declined during ovary maturation in adult female fish. These results suggest that the vasa gene play an important role in spermatogenesis and early oogenesis during the reproductive cycle of Korean rockfish.展开更多
目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快...目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快速扩增法（RACE）克隆其全长cDNA，应用生物信息学对其功能进行初步分析，应用一步法半定量RT—PCR检测其在AML患者及正常人中的表达。结果获得了家族性AML相关新基因FAMLF的cDNA全长，该基因定位于染色体1q31．3，cDNA全长2313bp，开放读码框（ORF）为249bp，编码82个氨基酸的蛋白质，含有信号肽，富含亮氨酸重复单位（LRR—SD22），内在固有无序结构域等功能区。Blast检索为功能未知的新基因，已被GenBank收录，并命名为FAMLF，核酸注册号为EF413001，蛋白质注册号为ABN58747。新基因FAMLF在AML患者中的表达明显高于正常人，差异有统计学意义（2．61±0．66 vs 0．97±0．51，P〈0．01）。结论成功获得一个家族性AML相关新基因FAMLF cDNA全长，FAMLF基因在AML中高表达，可能是具有一定生物功能的新基因。展开更多
In most bacteria, plants and algae, fatty acid biosynthesis is catalyzed by a group of freely dissociable proteins known as the type II fatty acid synthase （FAS II） system. In the FAS II system, enoyl- acyl carrier ...In most bacteria, plants and algae, fatty acid biosynthesis is catalyzed by a group of freely dissociable proteins known as the type II fatty acid synthase （FAS II） system. In the FAS II system, enoyl- acyl carrier protein reductase （ENR） acts as a determinant for completing the cycles of fatty acid elongation. In this study, the cDNA sequence of ENR, designated as IgENR, was isolated from the microalga lsochrysis galbana CCMM5001. RACE （rapid amplification of cDNA ends） was used to isolate the full-length cDNA oflgENR （1 503 bp）, which contains an open reading frame （ORF） of 1 044 bp and encodes a protein of 347 amino acids. The genomic DNA sequence oflgENR is interrupted by four introns. The putative amino acid sequence is homologous to the ENRs of seed plants and algae, and they contain common coenzyme- binding sites and active site motifs. Under different stress conditions, real-time quantitative polymerase chain reaction （RT-qPCR） showed the expression oflgENR was upregulated by high temperature （35℃）, and downregulated by depleted nitrogen （0 mol/L）. To clarify the mechanism of lipids accumulating lipids, other genes involved in lipids accumulation should be studied.展开更多
基金supported by The National Natural Science Foundation of China (No.41176122)the Key Program of Natural Science Foundation of Shandong,China (No.Z2008D03)
文摘Vasa, which is a conserved member of the DEAD-box protein family, plays an indispensable role in primordial germ cell proliferation. However, the expression ofvasa gene during the reproductive cycle in ovoviviparous fish has not been documented. In this study, the full-length sequence of vasa was obtained from the ovary of Korean rockfish （Sebastes schlegeli） using reverse transcription-PCR and rapid amplification of cDNA ends. The Vasa with a mature protein of 650 amino acids showed greatest homology （84%） with giant gourami （Osphronemus gorarny） and Pacific bluefin tuna （Thunnus orientalis）. The expression of vasa mRNA in Korean rockfish was detected in gonads only, suggesting its specific role in gonadal development. In addition, seasonal changes in the vasa expression levels were examined in gonads by quantitative real-time PCR. The vasa transcript levels in adult testis were found higher during spermatogenesis than during spermiation. The vasa transcript levels remained relatively high at the early ovary stage but declined during ovary maturation in adult female fish. These results suggest that the vasa gene play an important role in spermatogenesis and early oogenesis during the reproductive cycle of Korean rockfish.
文摘目的克隆家族性急性髓系白血病（AML）相关新基因cDNA全长，在分子水平上探讨急性白血病发生发展的机制。方法以构建的家族性AML抑制性消减文库中1个有差异表达的新基因EST序列zywb87（GenBank注册号CV973101）为基础，应用cDNA末端快速扩增法（RACE）克隆其全长cDNA，应用生物信息学对其功能进行初步分析，应用一步法半定量RT—PCR检测其在AML患者及正常人中的表达。结果获得了家族性AML相关新基因FAMLF的cDNA全长，该基因定位于染色体1q31．3，cDNA全长2313bp，开放读码框（ORF）为249bp，编码82个氨基酸的蛋白质，含有信号肽，富含亮氨酸重复单位（LRR—SD22），内在固有无序结构域等功能区。Blast检索为功能未知的新基因，已被GenBank收录，并命名为FAMLF，核酸注册号为EF413001，蛋白质注册号为ABN58747。新基因FAMLF在AML患者中的表达明显高于正常人，差异有统计学意义（2．61±0．66 vs 0．97±0．51，P〈0．01）。结论成功获得一个家族性AML相关新基因FAMLF cDNA全长，FAMLF基因在AML中高表达，可能是具有一定生物功能的新基因。
基金Supported by the National Natural Science Foundation of China(No.41106148)the Ocean Public Welfare Scientific Research Project(Nos.GHME2001SW02,200905019,200805039)the Science and Technology Development Program of Shandong Province(No.2011GHY11533)
文摘In most bacteria, plants and algae, fatty acid biosynthesis is catalyzed by a group of freely dissociable proteins known as the type II fatty acid synthase （FAS II） system. In the FAS II system, enoyl- acyl carrier protein reductase （ENR） acts as a determinant for completing the cycles of fatty acid elongation. In this study, the cDNA sequence of ENR, designated as IgENR, was isolated from the microalga lsochrysis galbana CCMM5001. RACE （rapid amplification of cDNA ends） was used to isolate the full-length cDNA oflgENR （1 503 bp）, which contains an open reading frame （ORF） of 1 044 bp and encodes a protein of 347 amino acids. The genomic DNA sequence oflgENR is interrupted by four introns. The putative amino acid sequence is homologous to the ENRs of seed plants and algae, and they contain common coenzyme- binding sites and active site motifs. Under different stress conditions, real-time quantitative polymerase chain reaction （RT-qPCR） showed the expression oflgENR was upregulated by high temperature （35℃）, and downregulated by depleted nitrogen （0 mol/L）. To clarify the mechanism of lipids accumulating lipids, other genes involved in lipids accumulation should be studied.
