南开大学学报(自然科学版)
南開大學學報(自然科學版)
남개대학학보(자연과학판)
ACTA SCIENTIARUM NATURALIUM UNIVERSITATIS NANKAIENSIS
2004年
1期
69-74
,共6页
刘翠敏%陶汉林%熊延%王宁宁%王淑芳%王勇
劉翠敏%陶漢林%熊延%王寧寧%王淑芳%王勇
류취민%도한림%웅연%왕저저%왕숙방%왕용
衰老%基因表达%丝氨酸-乙醛酸氨基转移酶%紫背浮萍
衰老%基因錶達%絲氨痠-乙醛痠氨基轉移酶%紫揹浮萍
쇠로%기인표체%사안산-을철산안기전이매%자배부평
senescence%gene expression%serine-glyoxylate aminotransferase%Spirodela polyrrhiza
本文报道对一个衰老下调基因的鉴定.从绿色和正在衰老的紫萍叶状体组织中分别提取总RNA,然后使用一个oligo-dT引物进行逆转录合成cDNA.cDNA用oligo-dT和简并引物进行PCR扩增,然后进行琼脂糖凝胶电泳.电泳后发现,一条eDNA带存在于由绿色组织而不存在于由正在衰老的组织所得到的样品中.将这一cDNA回收,克隆进T-vector并且进行了测序.Northern blot分析表明,该eDNA对应基因的表达受衰老调节而减少.根据第一个eDNA序列设计了一个特异性引物,进行RT-PCR扩增,并且克隆到另外一个部分eDNA.测序结果表明,第二个克隆具有与第一个eDNA相重叠的40bp的片段.将第一个eDNA序列与删去40bp重叠区的第二个eDNA序列连接,我们得到一个322个核苷酸的eDNA 3′端部分序列.用BLASTN程序将该序列翻译后与NCBI non-redundant database中已知的蛋白进行比较,结果表明所克隆的eDNA代表的基因与光呼吸途径的丝氨酸-乙醛酸氨基转移酶编码基因有很高的相似性.
本文報道對一箇衰老下調基因的鑒定.從綠色和正在衰老的紫萍葉狀體組織中分彆提取總RNA,然後使用一箇oligo-dT引物進行逆轉錄閤成cDNA.cDNA用oligo-dT和簡併引物進行PCR擴增,然後進行瓊脂糖凝膠電泳.電泳後髮現,一條eDNA帶存在于由綠色組織而不存在于由正在衰老的組織所得到的樣品中.將這一cDNA迴收,剋隆進T-vector併且進行瞭測序.Northern blot分析錶明,該eDNA對應基因的錶達受衰老調節而減少.根據第一箇eDNA序列設計瞭一箇特異性引物,進行RT-PCR擴增,併且剋隆到另外一箇部分eDNA.測序結果錶明,第二箇剋隆具有與第一箇eDNA相重疊的40bp的片段.將第一箇eDNA序列與刪去40bp重疊區的第二箇eDNA序列連接,我們得到一箇322箇覈苷痠的eDNA 3′耑部分序列.用BLASTN程序將該序列翻譯後與NCBI non-redundant database中已知的蛋白進行比較,結果錶明所剋隆的eDNA代錶的基因與光呼吸途徑的絲氨痠-乙醛痠氨基轉移酶編碼基因有很高的相似性.
본문보도대일개쇠로하조기인적감정.종록색화정재쇠로적자평협상체조직중분별제취총RNA,연후사용일개oligo-dT인물진행역전록합성cDNA.cDNA용oligo-dT화간병인물진행PCR확증,연후진행경지당응효전영.전영후발현,일조eDNA대존재우유록색조직이불존재우유정재쇠로적조직소득도적양품중.장저일cDNA회수,극륭진T-vector병차진행료측서.Northern blot분석표명,해eDNA대응기인적표체수쇠로조절이감소.근거제일개eDNA서렬설계료일개특이성인물,진행RT-PCR확증,병차극륭도령외일개부분eDNA.측서결과표명,제이개극륭구유여제일개eDNA상중첩적40bp적편단.장제일개eDNA서렬여산거40bp중첩구적제이개eDNA서렬련접,아문득도일개322개핵감산적eDNA 3′단부분서렬.용BLASTN정서장해서렬번역후여NCBI non-redundant database중이지적단백진행비교,결과표명소극륭적eDNA대표적기인여광호흡도경적사안산-을철산안기전이매편마기인유흔고적상사성.
The priliminary identification of a gene down-regulated by senescence is reported in this paper.Total RNA was isolated from green and senescing duckweed frond tissues and reverse-transcribed into cDNAsusing an oligo-dT primer. After amplification with PCR using the oligo-dT primer and degenerate primers, thecDNAs were electrophoresed on an agarose gel. One eDNA band was found to be present in the sample preparedfrom green but not from senescing tissues. This eDNA was recovered, cloned into a T-vector and sequenced.Northern blot analysis showed that the expression of the gene corresponding to the eDNA was down-regulatedby senescence. Another partial eDNA was also amplified by RT-PCR using a primer designed from the firstclone. Sequencing of the second clone shows that it contains an overlap part of 40bp with the first eDNA. Bymelding these two sequences we got a 3′ partial eDNA sequence of 322 nucleotides. This sequence wastranslated and compared with known proteins in the NCBI non-redundant database using the BLASTN program.The result shows that the gene corresponding to the cDNAs cloned has a very high similarity to genes encoding serine-glyoxylate aminotransferase, an enzyme involved in photorespiration.
