pUASTattB Drosophila Gen請紙e Expression Vector
(User-defined promoter)

Our pUASTattB ve照樹ctor system is 很筆highly effective for generati北那ng transgenic fl間大ies and controlling 身窗transgene expression. This system ut到水ilizes bacteriophage日民 φC31 integrase-mediated recombination 可一for efficient, targeted湖制 gene insertion, and can be used f暗店or achieving ub友暗iquitous, tissue-specific or i林影nducible transgene expression. T亮習he pUASTattB vector火錢 system is simila文麗r to the pUAST system, however 商廠it utilizes a φC31 integrase 美慢mediated recombination event rat時時her than P-element based transpositio雜拍n for genomic inser會坐tion of your gene of interest.

The bacteriophage φC31 encodes an 報劇integrase that 麗錯mediates efficient, sequence-s兒分pecific recombinati數又on between phage化在 attachment sites (called attP) and 事理bacterial attachment sites (cal數月led attB). In contra你數st to transposon-based喝就 systems, such as P-element-medi內見ated recombination, φC31-me電現diated insertion is i大女rreversible. Integration of物做 the pUASTattB into an attP site c歌議reates hybrid sites (called attL a市小nd attR), which are refractory算姐 to the φC31 integrase. Additionally務員, φC31-based insertion is site鐘家-specific, generally occurring only 了民at attP sites, and not話放 elsewhere in the geno答事me. For this reaso件你n, the pUASTattB vector system is des分風igned to be used with Drosophi市銀la lines carrying attP “landi但媽ng sites” within their genome.

The pUASTattB system cons友議ists of two vectors, both engine錢白ered as E. coli plasmids. One vector務離 referred to as the pUASTattB vector對媽 or the φC31 donor vecto睡唱r carries the attB 美新site and the user’s gene o低購f interest. The other vector re火河ferred to as the helper plasmid e木都ncodes for the φC31 低筆integrase. When 議醫the pUASTattB and the φC31 helper pla雪如smids are co-inject坐上ed into cells containi長影ng attP landing sites, φC31 i哥劇ntegrase-mediated recom熱腦bination between attB and attP 行老sites, results in the li區件nearization and int購熱egration of the pUASTatt兵船B vector into the 劇時host genome. The mini white ge飛畫ne on the pUASTattB vecto木煙r encodes for eye color and 子妹acts as a marker for the identific新費ation of transgenic flies which月但 have undergone s商月uccessful genetic recomb媽話ination of the transgene間話.

There are several possible metho商吧ds for introduction of the φC31 in制農tegrase into Drosop器海hila hosts. Target cells can be inject報從ed with φC31 integrase mRNA generated 錯睡by in vitro transc厭章ription. Alternatively, a hel我懂per plasmid expressing φC31 機秒integrase may be co-in離讀jected into the cells along with t月匠he pUASTattB vector.業坐 Additionally, there are Drosophila li高光nes available with germline 我南expression of φC31 inte子什grase which allow highly efficient tr音從ansgene insertion.

The user-defined promoter 行志version of the pUASTattB 路算Drosophila gene 年可expression vector&n話內bsp;allows users to select 年可a promoter of their choi是員ce from our Drosophila promoter d對村atabase for driving the expression of t影輛heir gene of interest (GOI) d答冷epending upon their e西呢xperimental goal. Our Drosophi麗樂la promoter datab銀見ase offers the following promoter家友 choices: ubiquitous promoters includ長綠ing actin 5C, polyubiqu看照itin and alpha-1 還資tubulin for driving ubiquitous ex看資pression of the GOI; tissue-specifi山遠c promoters such as R女這h2 for driving GOI expressi對靜on, specifically in Drosophila oc放務elli and inducible路一 promoters such as Mtn for achieving 少術inducible expression of the GOI with t件間he presence of Cu+.

For further information about this 購什vector system, please refer to 那拿the papers below.

Our pUASTattB Drosophila gene expressi得紙on vector is designed那白 to achieve eff草技icient φC31 integra術廠se-mediated genomic insertion&n兒作bsp;of a GOI. Our vectors嗎們 are optimized for hig廠哥h copy number replication in E. col體業i and high effic知畫iency transgenesis of 妹白Drosophila lines. The user-de和身fined promoter versi快歌on of this vector allo煙術ws users to select a ubiquit可裡ous, tissue-specific or 北雜inducible promoter for driving南場 their GOI depending upon their expe睡用rimental goal.

Technical complexity: The generation 還章of transgenic Drosoph嗎頻ila requires emb國聽ryonic injection and fly husb海技andry, which ca分鐘n be technically d藍新ifficult.

Requires attP insertion sites:&兒吃nbsp;The generation of tra子河nsgenic Drosophila using the p村東UASTattB vector s場鄉ystem requires th做空e use of specialized host件音 lines carrying attP “l東員anding sites” in their genome.