农产品加工·学刊(中)
農產品加工·學刊(中)
농산품가공·학간(중)
AEM RODUCTS ROCESSING
2014年
5期
19-20
,共2页
杨灵%徐慧琴%张志芳%李轶女
楊靈%徐慧琴%張誌芳%李軼女
양령%서혜금%장지방%리질녀
α-淀粉酶基因amyP%家蚕杆状病毒表达系统%重组杆状病毒
α-澱粉酶基因amyP%傢蠶桿狀病毒錶達繫統%重組桿狀病毒
α-정분매기인amyP%가잠간상병독표체계통%중조간상병독
α-amylase gene (amyP)%silkworm-baculovirus expression system%recombinant baculovirus
从高温菌Geobacillus. sp. POT5基因组中扩增得到推定的α-淀粉酶基因amyP,将其插入杆状病毒转移载体pVL1393上,获得重组转移载体pVL-amyP,经与线性化的亲本病毒DNA共转染家蚕细胞Bm-5后,通过体内同源重组的方式获得重组杆状病毒,将重组病毒感染家蚕5龄幼虫,提取发病家蚕血淋巴液进行淀粉酶活力分析,结果表明amyP基因在家蚕生物反应器中得到了正确的表达。
從高溫菌Geobacillus. sp. POT5基因組中擴增得到推定的α-澱粉酶基因amyP,將其插入桿狀病毒轉移載體pVL1393上,穫得重組轉移載體pVL-amyP,經與線性化的親本病毒DNA共轉染傢蠶細胞Bm-5後,通過體內同源重組的方式穫得重組桿狀病毒,將重組病毒感染傢蠶5齡幼蟲,提取髮病傢蠶血淋巴液進行澱粉酶活力分析,結果錶明amyP基因在傢蠶生物反應器中得到瞭正確的錶達。
종고온균Geobacillus. sp. POT5기인조중확증득도추정적α-정분매기인amyP,장기삽입간상병독전이재체pVL1393상,획득중조전이재체pVL-amyP,경여선성화적친본병독DNA공전염가잠세포Bm-5후,통과체내동원중조적방식획득중조간상병독,장중조병독감염가잠5령유충,제취발병가잠혈림파액진행정분매활력분석,결과표명amyP기인재가잠생물반응기중득도료정학적표체。
The putatived α-amylased gene is amplified from the genome of thermostable strain Geobacillus. sp. POT5 and is inserted into the transfer vector pVL1393. Thus the recombinant transfer vector pVL-amyP is obtained and co-transfected with linerizated parental virus DNA into the Bm-5 cells. The recombinant baculovirus is obtained by homologous recombination. Recombinant virus infect 5 instar larvae of silkworm, hemolymph is extracted and performed with activity analysis ofα-amylase. The results indicat that the amyP gene is correctly expressed in the silkworm.