| 1. | The polyhedrin gene sequence in recombinant virus bmpak - hbmp can be more benefit to enhancing the expression of the hbsag ( pres2 + s ) from atg initial codon at the 5 " ends of pres2 gene
另一方面， elisa检测表明， presz抗原性提高率约是hbsag的14倍，提示bmpak hb帅中多角体基因部分碱基序列的存在更有利于从presz前的atg开始表达中蛋白全基因（ presz s ） 。 |
| 2. | A 12 bp sequence of the 5 " end from the polyhedrin protein gene of bmnpv was ligated to the 5 ' end of hbsag ( pres2 + s ) protein coding sequence by pcr . the fusion product coding for hbv surface antigen medium sized ( hbmp ) with 4 - additional aa of bmnpv polyhedrin protein was obtained
本研究通过pcr突变的方法，在hbsag ( pres2 + s )前s2序列的5 ’端融合了bmnpv多角体蛋白基因5 ’端的12个碱基，获得了融合乙肝表面抗原中蛋白基因( hbmp ) 。 |
| 3. | To obtain large amounts of appa phytase , the appa gene was subcloned into the prokaryotic expression vector pet - 28a ( + ) and baculovirus transfer vector pvl - 1393 under the control of the lac and polyhedrin promoter , respectively . the appa phytase was overexpressed in e . coli strain bl21 induced by lactose
为了大量表达appa植酸酶，我们将appa基因分别克隆至原核表达载体pet - 28a ( + )和杆状病毒转移载体pvl - 1393中，将其分别置于lac和polyhedrin启动子控制之下。 |
| 4. | Hegf gene with his - tag at the end , which was derived from pet22 - egf , was in - frame fused to the carboxy - terminal of polyhedrin ( ph ) gene , which included the amino - terminal 116aa coding region . the polyhedrin - egf fusion gene ( named ph - egf ) was then cut out with ecorv and ecori , and was cloned between ecorv and ecori sites of pbacpak . 8 ( the result plasmid was named pbacph - egf and the ph - egf fusing gene was right under the control of ph promoter ) . the pbacph - egf structure was verified with restriction enzymes digestion and pcr
从pet22 - egf质粒中分离出末端带his - tag的egf基因，对位融合于多角体蛋白n端116个氨基酸基因序列的下游(命名为ph - egf ) ，并在两段基因间设计了凝血酶xa蛋白酶切位点，经过酶切、测序等鉴定正确后，克隆至pbacpak8中，使ph - egf融合基因置于多角体蛋白( polyhedrin ， ph )基因启动子控制之下，构建成重组转移载体pbacph - egf 。 |
| 5. | Two types of repeat sequence , a 15 - amino - acid ( eelcaqlcstppppi ) with 2 repeats and a 6 - amino - acid ( ppictp ) with 4 repeats , were firstly reported . 2 . the characterization of meq gene product and its expression within the cells a recombinant baculovirus transfer vector pblubac4 - meq was constructed by cloning meq gene of marek ' s disease virus ( mdv ) ga strain into the baculovirus transfer vector pbluebac4 under the polyhedrin promoter
此外，研究还发现了meq基因的两类有趣的重复结构，其中一类是含15个氨基酸残基( eelcaqlcstppppi )的结构，有2个重复，另一类是含6个氨基酸残基( ppictp )的结构，共有4个重复，它们全部分布在meq蛋白c -端的转录激活域内。 |
| 6. | To solve this problem , we used bac - to - bac system to recombine gfp - actin fusion gene and gfp gene under the control of the promoter of polyhedrin gene respectively . two recombinant virus : acmnpv - gfp - actin which contains gfp - actin fusion gene and acmnpv - gfp which contains gfp gene were obtained . the latter was set as a control
为了解决以上问题，在以上实验的基础上，我们利用bac - to - bac系统分别将gfp基因和gfp - actin融合基因重组于多角体基因启动子之下，构建了两种重组病毒，即含gfp - actin融合基因的重组病毒acmnpv - gfp - actin和含gfp基因的重组病毒acmnpv - gfp 。 |
| 7. | In the experiments discussed in chapter 5 we generated two recombinant viruses based on an acmnpv - and hasnpv - bac - to - bac system , respectively . in such recombinant viruses the busuctl gene under polyhedrin promoter was inserted into polyhedrin gene locus . a preliminary bioassay was conducted
第五章利用杆状病毒bac - to - bac系统构建了含有油桐尺蠖核多角体病毒的类蜗牛毒素基因的重组病毒racbacctl和rhabacctl ，在其相应宿主甜菜夜蛾和棉铃虫的细胞水平和虫体水平进行了超表达实验。 |
