Perera O P, Cockburn A F, Mitchell S E, Conn J, Seawright J A
Center for Medical, Agricultural, and Veterinary Entomology, Gainesville, Florida 32608, USA.
Am J Trop Med Hyg. 1998 Nov;59(5):673-8. doi: 10.4269/ajtmh.1998.59.673.
A genomic DNA library of Anopheles aquasalis Curry was screened for clones that hybridized more intensely to DNA from A. aquasalis than to DNA from A. benarrochi Gabaldon, Cova Garcia, and Lopez, A. konderi Galvao and Damasceno, A. nuneztovari Gabaldon cytotypes A, B, and C, A. oswaldoi (Peryassu), A. rangeli Gabaldon, Cova Garcia, and Lopez, or A. trinkae Faran. Two specific clones (2.5 kilobasepairs [kbp] and 3.0 kbp) from A. aquasalis were isolated. Both A. aquasalis-specific clones were from the intergenic spacer region of the ribosomal RNA (rRNA) cistron. Upon digestion with Rsa I, a 900-bp fragment from the clone AA-1 hybridized specifically to A. aquasalis DNA. Analysis of the DNA sequence of this fragment revealed four tandemly repeated 36-bp units. Three of these repeat units were identical, and the fourth was 94% identical to the others. The DNA sequence of a highly conserved region of these repeats was used to synthesize an oligonucleotide probe specific to A. aquasalis.
对库里按蚊(Anopheles aquasalis Curry)的基因组DNA文库进行筛选,寻找与库里按蚊DNA杂交信号比与贝氏按蚊(A. benarrochi Gabaldon、Cova Garcia和Lopez)、孔氏按蚊(A. konderi Galvao和Damasceno)、努氏按蚊(A. nuneztovari Gabaldon)细胞型A、B和C、奥斯瓦尔多按蚊(A. oswaldoi (Peryassu))、兰氏按蚊(A. rangeli Gabaldon、Cova Garcia和Lopez)或特氏按蚊(A. trinkae Faran)DNA杂交信号更强的克隆。从库里按蚊中分离出两个特定克隆(2.5千碱基对[kbp]和3.0 kbp)。这两个库里按蚊特异性克隆均来自核糖体RNA(rRNA)顺反子的基因间隔区。用Rsa I酶切后,克隆AA - 1的一个900 bp片段与库里按蚊DNA特异性杂交。对该片段的DNA序列分析揭示了四个串联重复的36 bp单元。其中三个重复单元相同,第四个与其他单元的同源性为94%。利用这些重复序列高度保守区域的DNA序列合成了一种库里按蚊特异性寡核苷酸探针。