Grossman G L, Campos Y, Severson D W, James A A
Department of Molecular Biology and Biochemistry, University of California, Irvine 92697-3900, USA.
Insect Biochem Mol Biol. 1997 Aug-Sep;27(8-9):769-81. doi: 10.1016/s0965-1748(97)00063-5.
Genomic DNA fragments encoding a salivary gland-specific alpha-amylase gene, Amylase I (Amy I), and an additional amylase, Amylase II (AmyII) of the yellow fever mosquito, Aedes aegypti, were isolated and characterized. Two independently isolated DNA fragments, G34-F and G34-14A, encode polymorphic alleles of Amy I. A 3.2 kilobase (kb) EcoR I fragment of G34-F, F2, has been sequenced in its entirety and contains 832 base pairs (bp) of the 5'-end, non-coding and putative promoter regions that are adjacent to 2.4 kb of the Amy I coding region. One intron, 59 bp in length, is found towards the 3'-end of the clone. A third genomic clone, 3A, corresponding to Amy II, was sequenced and shown not to contain the primary DNA sequence that encodes the 260 amino acid region that uniquely characterizes the amino terminal end of the Amy I product. Amy I was assigned by restriction fragment length polymorphism (RFLP) mapping to chromosome 2 (23.0 cM) and Amy II to chromosome 1 (44.0 cM). Amy I and Amy II are highly polymorphic and there may be multiple linked copies at each locus. Comparisons between Amy I and Amy II are presented for the putative promoter and conceptual translation products. The identification of two distinct amylase genes and their separate linkage assignments provides evidence for a multigene family of alpha-amylases in Ae. aegypti.
编码黄热病蚊子埃及伊蚊唾液腺特异性α-淀粉酶基因淀粉酶I(Amy I)和另一种淀粉酶淀粉酶II(AmyII)的基因组DNA片段被分离并进行了表征。两个独立分离的DNA片段G34-F和G34-14A编码Amy I的多态性等位基因。G34-F的一个3.2千碱基(kb)的EcoR I片段F2已被完整测序,包含5'端832个碱基对(bp)的非编码和推定启动子区域,该区域与Amy I编码区域的2.4 kb相邻。在克隆的3'端发现了一个长度为59 bp的内含子。对对应于Amy II的第三个基因组克隆3A进行了测序,结果表明其不包含编码Amy I产物独特的氨基末端260个氨基酸区域的主要DNA序列。通过限制性片段长度多态性(RFLP)图谱分析,Amy I被定位到2号染色体(23.0 cM),Amy II被定位到1号染色体(44.0 cM)。Amy I和Amy II具有高度多态性,每个位点可能有多个连锁拷贝。对推定启动子和概念性翻译产物进行了Amy I和Amy II之间的比较。两种不同淀粉酶基因的鉴定及其单独的连锁定位为埃及伊蚊α-淀粉酶多基因家族提供了证据。