pgem 中文意思是什麼

The total rna was isolated from pokeweed ( phytolacca americana ) leaves using the method of guanidine isothiocyante and used as template to amplify the total length and deleted mutant pokeweed antiviral protein ( pap ) gene by rt - pcr and then the pap gene was cloned into pgem - t vector. the sequencing results showed that pap gene had 99. 9 % identity comparing with the pap gene nucleotide sequence reported by lin et al ( 1991 ). the iptg - inducible expression vector containing the pap gene was constructed and transferred into e. coli bl21 ( de3 ) - plyss

將缺失型pap基因克隆到植物表達載體pbi121中，通過液氮冷凍法將重組質粒轉入農桿菌lba4404細胞中，然後採用葉盤法，在該農桿菌的介導下將pap基因導入普通煙草中，經過卡那黴素抗性篩選，最後獲得了轉pap基因的工程煙草植株，摩擦接種煙草花葉病毒（ tmv ） ，與非轉基因煙草相比，能夠推遲癥狀表現達2月之久，說明pap基因能夠在其它植物體內產生有活性的高抗病毒的蛋白質。

In our experiment, the specific fragment was amplified from transgenic bobwhite genome dna at annealing temperature 61 by using high - fidelity pfu dna polymerase and cloned into clone vector pgem - 7fz ( + ), then sequenced. the cloned sequence was completely identical to the sequence which was issued in genbank

本實驗採用了高保真pfudna聚合酶，在退火溫度61條件下從轉基因bobwhite品種基因組dna中擴增出特異性片段，將此片段插入克隆載體pgem - 7fz （ + ） ，經測序和序列分析表明，所擴增得到的片段含有bar基因完整的讀碼框，並且序列與genbank中發表的序列完全一致。

The 600 bp and 800 bp pcr products were cloned into the pgem - t easy vector. their cdna sequences were determined with positive clones or purified pcr product. conclusion : compared the 600 bp pcr product with the amino acids sequence for the fibrinolysin metalloproteinase from the venom of agkistrodon acutus from the southern of anhui province, their homology is 90. 6 %

結果：其中一對引物擴增得到一600bp產物；另一對引物擴增得到三條特異性的dna條帶，大小分別為1 . 5kb 、 1 . 3kb和800bp ，將600bp和800bpdna進行克隆及測序，並推導編碼的氨基酸序列。

In this paper, first strand cdna of 3abc gene was synthesized using template rna extracted from cells infected with fmdv. the complete 3abc gene about isoobp was amplified by pcr and ligated into pgem - t easy vector. after transforming e. coli dh5 a, ampicillin resistant colonies were isolated and plasmid dna was prepared and analyzed by restriction analysis and pcr. presence of the full length 3abc gene was verified by nucleotide sequence analysis and the plasmid containing the expected sequence was named as pgem - 3abc. comparing the aquired sequence of 3abc with that of reference strains, the homology is more than 99 percent. the pgem - 3abc was digested with sal i and bgl ii and ligated into xho i and bgl ii - digested expression vector ptriex - 4 neo. lt was identified by restriction analysis and pcr and sequencing that this fragment had a 17bp deletion hi the nucleotide sequence 708bp of 3abc gene, which happened to form a terminator codon behind 3ab gene, but it contained the complete open reading frame ( orf ) of 3ab gene. positive clones were selected and induced with lmmol / l isopropyl - d - galactoside ( iptg ), bacteria were detected by sds - page and western blotting after properly treated. the results showed that the 3ab gene expressed successfully in e. coli and 33. 5ku fusion protein can be recognized by the positive bovine serum of fmdv. the amount of target protein is over 26 % of the total bacteria protein by gel thin layer scanning analysis

擴增產物連接到pgem - teasy載體中，轉化大腸桿菌dh5菌株，篩選氨芐青霉素抗性菌落，提取質粒經酶切鑒定、 pcr分析以及確證性測序證明，所克隆的1500bp左右的片段含有完整的3abc基因，與國外參考序列相比，同源性在99以上。將重組質粒pgem - 3abc和表達載體ptriex - 4neo分別用sal和bgl與xho和bgl消化后，亞克隆3abc基因至原核表達載體ptriex - 4neo中，通過酶切鑒定、 pcr擴增以及序列分析，發現克隆到ptriex - 4neo載體上的片段於3abc基因708bp處出現了17bp的缺失，碰巧在3ab基因后形成一終止密碼子，但3ab基因的閱讀框架完整，選出含有3ab基因完整閱讀框架的陽性克隆，用iptg誘導表達，收集菌液進行sds - page電泳、 westernblotting分析，結果表明， 3ab基因在大腸桿菌中成功表達，其表達產物為分子量33 . 5ku的融合蛋白，並能被口蹄疫病毒陽性血清識別。經薄層掃描分析，表達量占總蛋白量的26以上。

Envelope gene gp85 of imc10200 subgroup j avian leukosis virus was cloned and expressed in the present study. the sequence encoding the gp85 domain of imc 10200 alv - j was amplified from pgem - imc2. 2 vector, which contains env gene of alv - j imc 10200 strain, and cloned into transfer vector pfast bacl

為深入探討alv - j的亞群特性，本研究利用alv - jgp85基因兩側的序列片段為引，物從正常spf蛋雞、商品肉雞和df1細胞基因組中完整地擴增了內源性類alv - jgp85基因。

4. the construction of middle - clone vector and expression vector the puc - cp and pgem - 7z plasmid were digested by kpnl and bamhi, and collected the digested cpti fragment and the pgem - 7z, then ligated by t4 dna ligase and formed the pgem - cp

中間載體及表達載體的構建將puc - cp質粒和pgem ? 7z質粒，用kpni和bamhi酶切，分別回收cpti片斷和酶切后的載體片段，用t _ 4連接酶連接構建成中間載體pgem - cp 。

Then the pcr product was purified, ligated into pgem - t vector by ta cloning

篩選陽性克隆，測序，大量制備序列完全正確的質粒。

Pgem - t vector system, jm109 competent cells, reverse transcription kit and in vitro translation kit were purchased from promega company

Pgem一t載體，感受態細菌jm109 ，反轉錄試劑盒和體外翻譯試劑盒均購自promega公司。

Clone human wnk4 full length cdna into pgem - t vector human kidney total una was extracted and used as template to amplify wnk4 full length cdna with the forward primer ( 7 - 27 ) and reverse primer ( 3808 - 3833 ) with the long template expanded pcr system kit

5 .原位雜交以小鼠腎臟總翩a為模板，擴增wnki基因外顯子1 ， 24一28 ，以a片段和wnk4基因外顯子13一16片段，純化后連接在pgem一t載體上。

In this article, the author amplified dna sequences encoding the mature peptides of human intact and truncated igf - 1 gene from human genomic dna by the new way of alternative pcr established and named as exons - piecing together method by this laboratory. the igf - 1 and des ( l - 3 ) igf - l gene were cloned in pgem - t vector and then subcloned in expression plasmid pgex - 3x. two recombinant constructs known as pgex - 3x - igf - l and pgex - 3x - des ( l - 3 ) igf - l were transformed to e. coli dh5 a respectively

我們利用外顯子拼接法從人的基因組dna中擴增了編碼完整型及截短型人igf - 1基因成熟肽的dna片段，並將其克隆至pgem - t載體中；經測序證明正確后，又構建了表達載體pgex - 3x - igf - 1及pgex - 3x - des （ 1 - 3 ） igf - 1 ，在大腸桿菌中進行了表達研究。

To make the pcr primer that have adequacy enzyme sit flowed the sequence, ligate with the pgem - t easy after pcr. make sure the sequence by sequencing

經組織培養后，以pcr和rt - pcr法檢測再生煙草，共得到轉synnhap基因煙草4株， pbi121空載體浸染的對照煙草再生苗5株。

To broaden our knowledge about regulatory molecules involved in stress response, we cloned the dreb1c from arabidopsis, and characterized its salt tolerance in transgenic research first. the flowing results were obtained : 1. the dreb1c full length was cloned from arabidopsis genome by pcr, and was inserted into pgem - t - easy vector

為了拓寬對參與植物脅迫應答的調控分子的認識，本論文從dreb1c的克隆著手，以擬南芥為材料，通過轉基因的方法，首次研究了dreb1c這種erebp / ap2類蛋白對植物耐鹽性的的影響，研究結果如下： 1

The total rna was purified from the germ in the liquid by the guanidine isothiocyantehod method, then the total rna digested by dnase that had not rnase was used for rt - pcr. i change the magnesium ion dencity in the pcr system in order to optimize the pcr condition. at the end i selected the magnesium ion density as 1. 25 mm. the production of rt - pcr was inserted directionally into pgem ? z ( ampr ). the pgem ? z ( ampr ) was used to transform e coli jm109. i got a positive clone through culling and identificatin. the dna sequence inserted into pgem ? z ( ampr ) was sequenced and blasted with the cdna sequence of the # - mannanase mature peptide that got from genbank

分取誘導培養液中的菌體，用異硫氰酸胍法提取總rna ，總rna再經無rna酶的dna酶處理後用于rt ? pcr 。在pcr擴增目的基因時，通過優選擴增體系，使鎂離子濃度為1 . 25mm時rt ? pcr可順利地獲得目的基因，並能定向克隆到載體pgem ? 3z （ amp ~ r ）中。用克隆載體轉化宿主大腸桿菌jm109 ，通過篩選獲取陽性克隆子，對陽性克隆子進行酶切與pcr鑒定，並對載體中插入的目的基因進行測序。

The results demonstrated that the momps were protective antigens and the momp - iscoms of aeromonas hydrophila could induce the host to mount satisfied immunity. a pair of primers were designed according to the published nucleotide sequence of a putative outer membrane protein gene ( omp ) of aeromonas hydrophila. with the specific primers, a target fragment about 1. 1kb was amplified from aeromonas hydrophila l316 via pcr. the target fragment was inserted into the linearized pgem - t easy vector

根據已發表嗜水氣單胞菌的外膜蛋白基因omp的核苷酸序列設計引物，利用pcr技術，擴增、克隆了嗜水氣單胞菌l316的主要外膜蛋白基因（ momp ） ，經t a克隆，插入到pgem - t系列載體上，測序分析結果表明momp基因最長的開放閱讀框（ orf ）為1035nt ，編碼由344個氨基酸組成，分子量為36kda的主要外膜蛋白質（ momp ） 。

To prepare the wild type mbl in prokaryotic system, a pair of primers was designed and synthesized, and was used to amplify mbl gene from the recombinant vector pgem - mbl that contans wild type mbl cdna. a recombinant prokaryotic expression vector, pet28 - mbl, was constructed by inserted the mbl gene into plasmid pet28 ( b ), and after transfected it into ecoli bl21 ( de3 ) and induced with iptg, recombinant mbl protein was expressed successfully

本實驗另選用了原核表達質粒pet28 （ b ） ，根據已構建好的含有mbl野生型基因的t載體pgem - mbl ，設計一對引物， pcr擴增mbl基因，凝膠回收，雙酶切pcr產物和pet28 （ b ）質粒， t4連接酶連接，轉化大腸桿菌dhsa ，氨芋選擇培養挑取克隆鑒定。

The amplified dna fragments were inserted into pgem - t easy vector and sequenced. the dna fragment sequencing results from the two subspecies were compared to detect whether there was any difference

將pcr擴增產物克隆到pgem - teasy載體，進行dna序列分析，並用生物信息學方法比較東方田鼠長江亞種與指名亞種之間該序列的差異。

On the bases of designing a primer pair, we obtained the coding domain sequence of rbp by pcr from cdna library. then the gene was cloned into pgem - t vector, the dna sequencing showed that the cloned gene was in agreement with the reported sequence. then the tageted gene of rbp was further subcloned into a procaryotic expression vector pbv220 and constructed recombinant plasmids was named pbv220 - rbp. in order to expresse rbp in procaryotic cell efficiently, the recombinant plasmids was introduced into e. colidhs a straints. expression of rbp was induced by temperature induction

本實驗在合成該蛋白基因上下游引物的基礎上，利用pcr技術，從人睪丸cdna庫中釣取目的基因，並克隆于pgem - t載體中，經序列測定證明與文獻報道基本一致，再將目的基因從重組克隆質粒中亞克隆于pbv220原核表達載體中，轉化宿主菌，經溫度誘導后，進行sds - page電泳分析，發現在約21kd位置上出現了一條明顯的蛋白帶，與預期相符。

Recombinant cloning vector of pgem - hc and pgem - ha were constructed by ligating the hc and ha genes into pgem t east vector and transformating into jm109. 3

分別將目的基因hypoderminc和hypodermina與克隆載體（ pgemteasy ）連接並轉化到宿主菌jm109中，構建了重組克隆載體pgem - hc和pgem - ha 。

These results showed that the three isolates were infectious bursal disease virus. the full - length cdna of the genomic segment a of two viruses, one virulent field strain ibdv zj2000 and one attenuated strains ibdv jd1, were amplified in a single step procedure by long - accurate reverse - transcription polymerase chain reaction ( la - pcr ), cloned into pgem - t easy vector, and sequenced

分別以傳染性法氏囊病病毒zj2000株（野毒株）和jd1株（弱毒株）基因組dsrna為模板，採用long - accuratert - pcr （ la - pcr ）一步法擴增並克隆了兩株病毒的基因組a節段全長cdna 。

Using recombination techniques, the gene fragment ced - 9, derived from pbs, was incorporated into pgem - 7zf ( + ) and formed a new vector pgem - ced9, in order to get the right endonuclease sites

從質粒pbs中獲取ced - 9基因片段，構建中間載體pgem - ced9以獲得合適的酶切位點。