红腺忍冬绿原酸生物合成新基因克隆及其生物信息学分析(1)
[摘要] 为挖掘红腺忍冬绿原酸生物合成的关键酶基因，应用生物信息学方法从红腺忍冬基因组数据库中筛选出4个新基因LHPAL4，LHHCT1，LHHCT2，LHHCT3，并预测其编码蛋白的结构和功能。序列分析结果表明，红腺忍冬LHPAL4与LHPAL1关系密切;LHHCT1，LHHCT3，LHHQT2关系密切，LHHCT2单独聚为一类。通过Real-time PCR 检测4个新基因在红腺忍冬不同组织中的转录情况，发现LHPAL4，LHHCT1，LHHCT3在败花中表达量最高，LHHCT2在叶中的表达量最高，为揭示不同部位活性成分含量差异机制提供依据，也为活性成分的合成生物学提供元件。
[关键词] 红腺忍冬;活性成分;基因家族;变异分析
[收稿日期] 2014-09-20
[基金项目] 国家自然科学基金项目(81373959);国家杰出青年科学基金项目(81325023)
, 百拇医药
[通信作者] 黄璐琦，E-mail：huangluqi01@126.com;袁媛，E-mail：yuanyuan@icmm.ac.cn
Gene cloning and bioinformatics analysis of new gene for chlorogenic acid
biosynthesis of Lonicera hypoglauca
YU Shu-lin， HUANG Lu-qi， YUAN Yuan， QI Lin-jie， LIU Da-hui
(1.Pharmacy College， Anhui University of Chinese Medicine， Hefei 230038， China;
2. State Key Laboratory of Dao-di Herbs， National Resource Center for Chinese Materia Medica， Academy of
, 百拇医药
Chinese Medical Sciences， Beijing 100700， China;
3. Institute of Medicinal Plant， Yunnan Academy of Agricultural Sciences， Kunming 650000， China)
[Abstract] To obtain the key genes for chlorogenic acid biosynthesis of Lonicera hypoglauca， four new genes ware obtained from the our dataset of L. hypoglauca. And we also predicted the structure and function of LHPAL4， LHHCT1， LHHCT2 and LHHCT3 proteins.The phylogenetic tree showed that LHPAL4 was closely related with LHPAL1， LHHCT1 was closely related with LHHCT3， LHHCT2 clustered into a single group. By Real-time PCR to detect the gene expressed level in different organs of L. hypoglauca， we found that the transcripted level of LHPAL4， LHHCT1 and LHHCT3 was the highest in defeat flowers， and the transcripted level of LHHCT2 was the highest in leaves. These result provided a basis to further analysis the mechanism of active ingredients in different organs， as well as the element for in vitro biosynthesis of active ingredients.
, 百拇医药
[Key words] Lonicera hypoglauca; active ingredients; gene family; variance analysis
doi：10.4268/cjcmm20150515
红腺忍冬Lonicera hypoglauca Miq.为忍冬科忍冬属植物，又名菰腺忍冬、盘腺忍冬、腺背忍冬等，其花蕾或初开的花被作为药材山银花使用[1]。红腺忍冬中的主要活性成分为酚酸类、黄酮类化合物，其中绿原酸(chlorogenic acid)的含量最高[2]，具有抗菌、抗病毒、增高白血球、保肝利胆、抗肿瘤、降血压、降血脂、清除自由基和兴奋中枢神经系统等作用[3]。绿原酸是由咖啡酸(caffeic acid)与奎尼酸(quinic acid)生成的缩酚酸，是植物体在有氧呼吸过程中经莽草酸途径产生的一种苯丙素类化合物[4]，其最初是由L-苯丙氨酸在苯丙氨酸解旋酶(phenylalanine amononiar-lyase，PAL)，4-coumarate CoA ligase (4CL)，cinnamate 4-hydroxylase (C4H)的作用下生成p-coumaroyl-CoA，然后在hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT)的作用下生成绿原酸[5]。, http://www.100md.com(余淑琳 黄璐琦 袁媛 齐琳洁 刘大会)
[关键词] 红腺忍冬;活性成分;基因家族;变异分析
[收稿日期] 2014-09-20
[基金项目] 国家自然科学基金项目(81373959);国家杰出青年科学基金项目(81325023)
, 百拇医药
[通信作者] 黄璐琦，E-mail：huangluqi01@126.com;袁媛，E-mail：yuanyuan@icmm.ac.cn
Gene cloning and bioinformatics analysis of new gene for chlorogenic acid
biosynthesis of Lonicera hypoglauca
YU Shu-lin， HUANG Lu-qi， YUAN Yuan， QI Lin-jie， LIU Da-hui
(1.Pharmacy College， Anhui University of Chinese Medicine， Hefei 230038， China;
2. State Key Laboratory of Dao-di Herbs， National Resource Center for Chinese Materia Medica， Academy of
, 百拇医药
Chinese Medical Sciences， Beijing 100700， China;
3. Institute of Medicinal Plant， Yunnan Academy of Agricultural Sciences， Kunming 650000， China)
[Abstract] To obtain the key genes for chlorogenic acid biosynthesis of Lonicera hypoglauca， four new genes ware obtained from the our dataset of L. hypoglauca. And we also predicted the structure and function of LHPAL4， LHHCT1， LHHCT2 and LHHCT3 proteins.The phylogenetic tree showed that LHPAL4 was closely related with LHPAL1， LHHCT1 was closely related with LHHCT3， LHHCT2 clustered into a single group. By Real-time PCR to detect the gene expressed level in different organs of L. hypoglauca， we found that the transcripted level of LHPAL4， LHHCT1 and LHHCT3 was the highest in defeat flowers， and the transcripted level of LHHCT2 was the highest in leaves. These result provided a basis to further analysis the mechanism of active ingredients in different organs， as well as the element for in vitro biosynthesis of active ingredients.
, 百拇医药
[Key words] Lonicera hypoglauca; active ingredients; gene family; variance analysis
doi：10.4268/cjcmm20150515
红腺忍冬Lonicera hypoglauca Miq.为忍冬科忍冬属植物，又名菰腺忍冬、盘腺忍冬、腺背忍冬等，其花蕾或初开的花被作为药材山银花使用[1]。红腺忍冬中的主要活性成分为酚酸类、黄酮类化合物，其中绿原酸(chlorogenic acid)的含量最高[2]，具有抗菌、抗病毒、增高白血球、保肝利胆、抗肿瘤、降血压、降血脂、清除自由基和兴奋中枢神经系统等作用[3]。绿原酸是由咖啡酸(caffeic acid)与奎尼酸(quinic acid)生成的缩酚酸，是植物体在有氧呼吸过程中经莽草酸途径产生的一种苯丙素类化合物[4]，其最初是由L-苯丙氨酸在苯丙氨酸解旋酶(phenylalanine amononiar-lyase，PAL)，4-coumarate CoA ligase (4CL)，cinnamate 4-hydroxylase (C4H)的作用下生成p-coumaroyl-CoA，然后在hydroxycinnamoyl-CoA quinate hydroxycinnamoyl transferase (HQT)的作用下生成绿原酸[5]。, http://www.100md.com(余淑琳 黄璐琦 袁媛 齐琳洁 刘大会)