CAJ | 학술논문

Kinesin家族是一类马达蛋白,它们能利用ATP水解所释放的能量驱动自身携带物质分子沿着微管运动,在细胞形成、细胞伸长等方面起着关键作用。本研究以拟南芥 Atkinesin-13A 蛋白序列作为探针序列,利用 Blast 比对从二倍体雷蒙德氏棉的基因组数据库中发现7个具有较高同源关系的基因。根据基因序列设计引物,利用 RT-P C R技术从陆地棉纤维中分离出7个基因。依据7个基因与Atkinesin-13A和Atkinesin-13B的同源性高低,依次将其命名为GhKIS13A1、GhKIS13A2、GhKIS13A3、GhKIS13B1、GhKIS13B2、GhKIS13B3和GhKIS13B4。生物信息学分析表明,7个Ghkinesin13均含有典型的KISC马达区域、ATP结合位点和微管结合位点,其马达区域属于中央马达。多重序列比对和进化树分析发现,这7个蛋白可被分为2个(Kinesin13A和Kinesin13B)亚类。实时荧光定量PCR结果表明,7个Ghkinesin13亚家族基因在棉花各组织中均有表达,但表达模式各不相同,其中GhKIS13B4在纤维中优势表达,表明其在纤维发育过程中可能发挥着重要作用。
Kinesin가족시일류마체단백,타문능이용ATP수해소석방적능량구동자신휴대물질분자연착미관운동,재세포형성、세포신장등방면기착관건작용。본연구이의남개 Atkinesin-13A 단백서렬작위탐침서렬,이용 Blast 비대종이배체뢰몽덕씨면적기인조수거고중발현7개구유교고동원관계적기인。근거기인서렬설계인물,이용 RT-P C R기술종륙지면섬유중분리출7개기인。의거7개기인여Atkinesin-13A화Atkinesin-13B적동원성고저,의차장기명명위GhKIS13A1、GhKIS13A2、GhKIS13A3、GhKIS13B1、GhKIS13B2、GhKIS13B3화GhKIS13B4。생물신식학분석표명,7개Ghkinesin13균함유전형적KISC마체구역、ATP결합위점화미관결합위점,기마체구역속우중앙마체。다중서렬비대화진화수분석발현,저7개단백가피분위2개(Kinesin13A화Kinesin13B)아류。실시형광정량PCR결과표명,7개Ghkinesin13아가족기인재면화각조직중균유표체,단표체모식각불상동,기중GhKIS13B4재섬유중우세표체,표명기재섬유발육과정중가능발휘착중요작용。
Kinesin family belongs to a class of motor proteins. Kinesins can move along microtubule filaments by using the en-ergy released from ATP hydrolysis, and play key roles during cell formation and cell elongation. Using the protein sequence of Atkinesin-13A as a probe, seven genes with high sequence homology were obtained from a genome database ofGossypium rai-mondiidiploid cotton with Blast alignment. These sequences were used to design primers, and then seven genes were isolated from upland cotton fiberusing RT-PCR. Based on the homology level of the seven genes withAtkinesin-13A andAtkinesin-13B, we designated them asGhKIS13A1, GhKIS13A2, GhKIS13A3, GhKIS13B1, GhKIS13B2, GhKIS13B3, and GhKIS13B4 respec-tively. Bioinformatic analysis showed that seven proteins contained typical KISC domains including the central motor, ATP- binding sites and microtubule binding sites. Multiple sequence alignment and phylogenetic tree analysis revealed that seven pro-teins can be divided into Kinesin13A and Kinesin13B. The qPCR showed that the sevenGhkinesin13 subfamily genes expressed in all tissues of cotton, but showed different expression patterns. Of the seven genes, onlyGhKIS13A4 was preferentially ex-pressed in fibers, suggesting that it may play an important role in cotton fiber development.