Cloning, Sequences Analysis And Seni-quantitative Expression Analysis Of The Apidermin Cuticular Protein Family From Apis Ceratta

cDNA sequences of six Apidermin cuticular protein genes from Apis ceranawere cloned in this study, structures and sequence features of the genes were anlysedwith bioinformatics approaches, and expression level of them were detected, thecharacteristics of insect Apidermin cuticular protein family were synthetically studiedat last. The results were summarized as follows:(1) A cluster of six apd genes were tandemly arranged in a GC-rich36kbgenomic region of A. cerana genome, and their length of cDNA sequences were:1478bp(apd-1),509bp(apd-2),524bp(apd-3),546bp(apd-like),1213bp(apd-1-like)and871bp(apd-3-like).(1) Six cuticular proteins of Apidermin family from Apis cerana are highlyhydrophobic and have extremely similar signal peptide sequences, severaltranscription factor binding sites of apidermin genes in the promoter sequences almostwere related to the process of insect metamorphosis reaction. Seventeen insectcuticular proteins of Apidermin family were Systematically Analyzed withbioinformatics approaches. We found the repetitive conservative sequence of12amino acid(GP-x(2)-GP-x(4)-GP) in the insect Apidermin family for the first time,sectional CPG family cuticular protein motifs were found also, and the Apiderminfamily genes were highly conservative between Apis cerana and Apis mellifera.(1)The expression level of six cuticular protein genes from Apis cerana worker atdifferent developmental stages(little larvae, big larvae, white pupae, dark pupae,newly emerging and forging) and with different tissues of the body(tentacle, poisonsac, muscle, midgut, brain) were detected by seni-quantitative RT-PCR technique, thisstudy showed that the apidermin genes from Apis cerana have specific andSpatio-temporal expression pattern. It shows expression signal at all stages beforeemerging period, the expreesion signal is weak on the periods of larvae and whitepupae, but the expression level is go to the top on period of newly emerging and dark pupae, and there is no expression signal on the period of forging. All the experimentalmeterials for RT-PCR seni-quantitative expression analysis with different tissues weredissected from newly emerging worker. apd-1is proved that expressed in theepidermal cell of Apis mellifera was detected expression signal only in the tissues oftentacle and poison sac. apd-2was not detected expression signal only in the tissuesof brain. apd-3was detected expression signal only in the tissues of tentacle, poisonsac and muscle. The genes including apd-1-like, apd-3-like and apd-like were detectedweak expression signal only in the tissues of tentacle, poison sac and muscle.